A preclinical approach to textile finishing and evaluation of bamboo fabric coated Acalypha indica and Tectona grandis for wound-dressing material

Abstract

This study explores the potential of Acalypha indica and Tectona grandis leaf extracts in wound-dressing applications, emphasizing the development of natural, bio-based treatment options. Using ultrasonic-assisted extraction (UAE) with ethanol, the extracts were coated onto bamboo woven fiber fabric at varying concentrations (50%, 75%, and 100%). Comprehensive characterization of bioactive compounds was conducted through phytochemical analysis, thin layer chromatography (TLC), antioxidant assays (FRAP), gas chromatography mass spectrometry (GC-MS), and Fourier-transform infrared (FTIR) spectroscopy. The coated fabric exhibited effective antibacterial properties against Bacillus subtilis and Escherichia coli. In addition, tests on fragrance, color intensity, and olfaction showed that the treated fabric maintained a pleasant herbal aroma lasting up to 20 washes. To further investigate the role of bamboo fabric as a substrate, an in vivo wound healing assay was performed on Wistar rats over 21 days. The 75% and 100% extract concentrations demonstrated over 90% wound closure, with the fastest healing time being 17 days. These results highlight that bio-extract-treated bamboo fabric not only promotes wound healing but also offers textile-related advantages, positioning it as a promising candidate for functional healthcare textiles.

Keywords

gas chromatography mass spectrometry (GC-MS),Fourier-transform infrared (FTIR) spectroscopy,Olfaction assay,antimicrobial activity,antioxidant activity,wound dressing,wound healing

The skin, the body’s largest sensory organ, acts as a crucial barrier against microbial, chemical, and physical assaults. When this barrier is compromised, as in wounds, the skin’s layered architecture is damaged, and a series of biological responses initiate the healing process.¹ Wounds can arise from various causes: burns, surgical procedures, accidents, and chronic conditions such as diabetes and sickle cell disease, which hinder the normal healing process and increase vulnerability to infection and prolonged recovery. Classifications of wounds, such as acute versus chronic and open versus closed, aid in understanding their treatment needs. Closed wounds, for example, restrict blood flow within the body, posing unique challenges to healing.² The skin’s outermost layer, the epidermis, is rich in protective features such as sebaceous and sweat glands, hair follicles, and an extracellular matrix that supplies resilience, immunity, and nutrient flow. Wound healing occurs in four phases, hemostasis, inflammation, growth, re-epithelialization, and remodeling, each essential for tissue repair and regeneration. However, complications arise in individuals with compromised healing processes, emphasizing the need for effective wound care solutions. The field of pharmacology has long recognized the therapeutic value of plants.³ Increasingly, plant-based medicines are favored for their low toxicity and minimal side effects, enhancing their appeal as alternatives to synthetic treatments. Secondary metabolites in plants, essential for growth, communication, and environmental protection, also offer potent healing and antibacterial properties, making them valuable in wound care.⁴

This study focuses on the wound-healing effects of Acalypha indica and Tectona grandis, two plants with rich medicinal histories, integrated into bamboo fabric for wound dressing. A. indica, often viewed as a common weed in tropical climates, is known for its use in traditional medicine across diverse cultures. Studies highlight its utility in treating ailments such as bronchitis, asthma, ulcers, and skin infections, demonstrating the plant’s potential for wound healing and antibacterial applications.⁵ Similarly, T. grandis (teak), a plant with traditional roots in the Verbenaceae family, offers anti-inflammatory, expectorant, and wound-healing properties. Teak extracts have long been utilized in folk medicine to treat ailments including bronchitis, hyperacidity, and various types of wounds, especially burns.⁶ The combined use of A. indica and T. grandis offers a promising plant-based approach to wound care.

Role of bamboo fabric in wound dressing

In wound care, dressings provide the first line of defense, arresting blood flow and creating an environment conducive to healing. The substrate material is pivotal, and bamboo fabric stands out for its unique properties: antibacterial activity, biodegradability, high moisture absorption, softness, and UV protection. Bamboo’s resilience to environmental conditions and extensive availability make it a valuable resource. In addition, the textile industry is increasingly exploring sustainable, eco-friendly materials such as bamboo fiber, which boasts high tensile strength, superior moisture absorption, and breathability, qualities advantageous for wound care and comfort.

Bamboo fabric has been shown to facilitate a moist, breathable environment ideal for wound-dressing applications.⁷ Its natural antibacterial properties, combined with the bioactivity of plant extracts such as A. indica and T. grandis, offer an innovative solution for enhancing wound healing efficacy. The fabric’s ability to retain the active compounds’ antibacterial and therapeutic properties through multiple washes adds further practicality to its application.

Current study and research significance

This study integrates bamboo fabric with the bioactive extracts of A. indica and T. grandis to develop a natural wound-dressing material. By leveraging the antibacterial and wound-healing properties of these plant extracts, alongside bamboo’s inherent benefits, we aim to create a functional textile for healthcare that aligns with sustainable practices in textile production.⁸ Furthermore, understanding the bamboo fabric’s role in supporting these bioactive compounds and enhancing their effects during wound healing presents a novel perspective for researchers and medical practitioners interested in plant-based, sustainable wound-care solutions.

Materials and methods

Plant collection

The leaves of A. indica and T. grandis for wound dressing were collected from local fields in Pollachi, Tamilnadu, India, and washed with running tap water. Washed leaves were then pat dried with clean cloth and shade dried at 37°C for 48 hours.

Ultrasonic-assisted extraction

The dried leaves were then made into fine powder which was suspended with ethanol (1:10) in order to hold the bioactive compounds, present in leaves. After this pretreatment, the powdered samples were subjected to ultrasonic-assisted extraction (UAE) utilizing ethanol–water (50:50) solution. The ultrasonic light analyses were completed in a LABMAN Probe sonicator (model PRO-650) working at 60-W power and 50-kHz recurrence. Measurements of the tank were 150 mm × 140 mm × 100 mm. Through this analyzer, desired compounds were retrieved by rupturing the cell wall using ultrasonic sound waves with optimum condition (pulse 5 s, temperature −5°C, time 30 min). The prepared extract was then exposed to dark maceration, where the sonicated extract is kept in dark for overnight by wrapping with foil and stored at room temperature for further analysis. The leaf extract is then separated and diluted at different concentrations of 100%, 75%, and 50%.⁹

Fabric finishing by bioactive extracts

The extract was coated on a fabric by a laboratory scaled padder. The pressure between the pads was set at 2 bars and the rotation speed set to 2 m/min. The fabric was incorporated with the extracts of various concentration and the wet fabrics were dried using hot air oven at 55°C for 20 min. Then the dried and coated fabrics were kept at atmospheric room temperature for 24 hours.¹⁰

Preliminary qualitative analysis

Test for tannins (ferric chloride test)

We added 5% of ferric chloride solution to 1 ml of extract. The appearance of blue–black indicates the presence of tannins.

Test for saponins (foam test)

The extract was shaken vigorously with 20 ml of water and observed for persistent foam which indicates the presence of saponins.

Test for flavonoids (alkaline reagent test)

A few drops of 10% NAOH was added to 1 ml of extract. An intense yellow color will appear which becomes colorless on addition of dilute acid which indicates the presence of flavonoids.

Test for alkaloids (Wagner test)

We dissolved 2 g of iodine and 6 g of potassium iodide in 10 ml of distilled water. We added 1 ml of extract to 1 ml of prepared reagent and reddish-brown color indicates the presence of alkaloids.

Test for cardiac glycosides (Keller–Killani test)

To the test tubes containing 1 ml of extract, 0.5 ml of glacial acetic acid, 2 drops of 5% ferric chloride, and concentrated sulfuric acid were added and the observed disappearance of reddish-brown color at the junction of two layers and bluish green in the upper layer indicate the presence of cardiac glycosides.

Test for terpenoids

To 1 ml of extract we added a few drops of concentrated H₂SO₄. The appearance of reddish brown color indicates the presence of terpenoids.

Test for steroids

To 5 ml of aqueous plant extract we added 2 ml of chloroform and concentrated H₂SO_4. The upper layer in the test tube turns red and the H₂SO₄ layer showing yellow with green fluorescence indicates the presence of steroids.¹¹

Thin layer chromatography

The phytoconstituents present in the organic extracts were determined qualitatively by thin layer chromatography (TLC). It was used to characterize the non-volatile mixtures of extract. In this, aluminum foil coated with silica gel on (60 F₂₅₄ plate) was used under various mobile phases for bioactive compound (secondary metabolite) identification. In TLC, the extracts spotted on silica coated plates, were analyzed for tannins, saponins, flavonoids, cardiac glycosides, terpenoids, steroids, and, finally, for alkaloids. The tannin contents were determined using chloroform:ethyl acetate:methanol (50:30:20). The characterization of saponin and flavonoid contents was resolved using chloroform:methanol:water (62:36:2)¹² and chloroform:ethanol:glacial acid (94:5:1).¹³ The alkaloids and cardiac glycosides were observed using ethyl acetate:methanol:water (81:11:8). Detection of terpenoid components was observed by using the mobile phase as toluene:ethyl acetate (93:7)¹⁴ and steroid components as chloroform:acetone (80:20).¹⁵ The developed plates were then sprayed with vanillin solution (1% w/v in 50% phosphoric acid) for steroid detection and Wagner’s reagent for alkaloid detection. The TLC results were further used to validate the presence of tannins based on positive reaction (brownish green–blue black coloration) with 0.1% FeCl₃, alkaloids based on positive reaction (brown coloration) with Wagner’s reagent, steroids based on positive reactions (violet to blue or green) with acetic anhydride and H₂SO₄, steroidal glycosides by Keller–Killani test, and cynogenic-glycoside-based red coloration of picrate paper.

Gas chromatography and mass spectrometry analysis

Gas chromatography (GC) was performed using the ethanolic extract of A. indica and T. grandis. The extract was analyzed using GC-mass spectrometry (MS; Perkin Elmer model: clarus 680) which is fortified with mass spectrometer clarus 600 (EI) and studied using Turbo Mass version 5.4.2 software. A fused silica column which is wrapped with Elite-5MS (5% biphenyl 95% dimethylpolysiloxane, 30 m × 0.25 mm ID × 250 µm df). Helium was employed as a carrier gas with a constant flow rate of 1 ml/min to separate the components. The temperature of the injector was set at 260°C when the chromatography began to run. Then 1 µl of sample was injected into the equipment and then oven was heated to 60°C for approximately 2 minutes, followed by 300°C at a rate of 10°C min⁻¹, and 300°C for about 6 minutes. The mass detector was operated under the following conditions: scan time of 0.2 seconds, scan interval of 0.1 seconds, ionization mode electron impact at 70 eV, transfer line temperature of 230°C, ion source temperature of 230°C, and ionization mode electron impact at 70 eV, a scan time 0.2 s, and scan interval of 0.1 s. The fragments appear from 40 to 600 Da. The spectrum of the components was compared with the database of known components stored in the GC-MS NIST (2008) library. The prominent compounds identified were within the retention time of 1.59.¹⁶

Fourier-transform infrared spectroscopy

The bio-extract powder of A. indica and T. grandis was subjected to Fourier-transform infrared (FTIR) spectroscopy for further validation of secondary structures. The samples were taken in the ratio of 1:1 (conc.) and it was mixed with KBr and it is pelletized by KBr hydraulic press of pressure of about 10 tons. FTIR spectroscopy was used to examine the functional parts in the pellet at the IR range of 700–4000 cm^–1.

Antioxidant assay

The reducing power of the extracts was determined by ferric reducing antioxidant power assay (FRAP). FRAP was performed with five different concentrations of Acalypha indica Tectona grandis extracts (0.2, 0.4, 0.6, 0.8, and 1 mg/ml) and L-ascorbic acid at the same concentrations, which is then mixed with 2 ml phosphate buffer after (0.2 M, pH 6.6) and 2 ml of 1% potassium ferricyanide (K₃Fe(CN)₆). The mixture was then incubated at 50°C for 20 minutes. Afterwards, 2 ml of 10% trichloroacetic acid (TCA) was added, and mixture was centrifuged at 3000 rpm (revolutions per minute) for 5 minutes. The supernatant (2 ml) was aspirated and mixed with 0.1% ferric chloride (FeCl₃) and 2 ml of distilled water. In every instance, the experiment was performed in triplicate. Then, the absorbance was measured spectrophotometrically at 700 nm using a UV–vis spectrophotometer and observed.¹⁷

Treatment of bamboo fabric with chitosan

The fabric was treated with 5% of chitosan. The material and liquor ratio is 1:30. The chitosan was dissolved in water and kept at a water bath temperature of 70°C. The fabric was treated for about 1 hour and dried in a hot air oven.

Antimicrobial finish application of bamboo fabric

The fabrics were absorbed in the 50%, 75%, and 100% concentrations of ethanolic extracted consortium of A. indica and T. grandis for 30 minutes and padded separately and compressed to get a wet pick up of 85% on the weight of the fabric. At 80°C for 3 min the fabric was dried and restored at 95°C for 10 min on a lab model curing chamber.

Antimicrobial activity assessment of dyed bamboo fabric

Antimicrobial analysis of bamboo fabric treated with A. indica and T. grandis was carried out using the nutrient agar plate method. On the petri dish, the nutrient agar medium was prepared. The gram-negative (Escherichia coli) and gram-positive bacteria (Bacillus subtilis) were used for the antimicrobial activity. The prepared nutrient agar plate was inoculated with the above bacterial culture using an L rod technique and the plates were kept uninterrupted for 24 hours at 37°C in an incubator. After 24 hours, the antibacterial activity was evaluated using qualitative assessment by the agar well diffusion method (ISO 20645). The dyed fabric that is fabric treated with A. indica and T. grandis and undyed control fabric samples were placed in close contact with AATCC bacteriostatic nutrient agar medium plate, the plates were re-incubated at 37°C for 18–24 h. After re-incubation, the zone of inhibition analysis was analyzed and calculated to interrupt the antibacterial efficiency of bio-consortia-treated fabric. The competency of antimicrobial action of A. indica and T. grandis on bamboo fabric is indicated by the zone formation.¹⁸

Determination of color strength

Samples treated with A. indica and T. grandis were tested in a reflectance spectrophotometer (Gretagmacbeth) with D65 light using Colour-i-control software joined with an IBM PC. The K/S value for comparison of color between bamboo samples treated with A. indica and T. grandis solution and washed samples treated with different cycles was evaluated using a Chroma flash Color Matching System (ASHCO Industries Ltd., India). The Kubelka–Munk equation is used to calculate the color value K/S:

\frac{K}{S} = \frac{{(1 - R)}^{2}}{2 R} \times 100

where K is the absorption coefficient, S is the scattering coefficient, and R is the spectral reflectance of the colored samples at a wavelength of maximum absorption (where the reflection is minimum).

A color fastness to washing test was performed according to standard ISO 105 C06 A2S using a solution of 5 g/l of soap in which free alkali calculated as Na₂CO₃ should not be more than 0.3%, free alkali calculated as NaOH should not be more than 0.1%, and the total fatty matter should not be more than 85%. Fabric samples were treated at 40 + 2°C for 30 minutes. The test was performed in the gyrowash machine. The treated samples are rinsed in cold distilled water following 10 minutes of rinsing in cold running tap water. After squeezing the fabric is dried in air at a temperature, not above 60°C. The change in color of the fabric is assessed with Gray Scale No. 1. The samples were further evaluated for fragrance.¹⁹

Fragrance intensity test

The life span of a fragrance is based on the concentration of the scent and is completely based on the fragrant compounds. The samples treated with A. indica and T. grandis were evaluated for fragrance intensity in a subjective test method wherein judges had given the rating on a five-point Likert scale, i.e., 4 stands for excellent, 3 for good, 2 for fair, 1 for poor, and 0 is for nil fragrance intensity. The experiment was carried out over 4 weeks.²⁰

Olfaction China GB test

This method was performed quickly after opening the test samples. The method was conducted in an environment that was free from unconventional scents. Once the tester had their washed hands and put on gloves, they took the fabric specimen close to the nostril and smelled the aroma in the fabric. Meanwhile, the aromatic hydrocarbons were traced and the unconventional scent was subsequently noted. The process was carried out by two personnel and the result was analyzed based on the existing standard code GB 18401-2010 which represents national general safety technical code for textile products (ChineseStandard.net 2010).

Experimental protocol for in vivo study conducted in wounded rats to validate the efficiency of the bio-extract-coated fabrics

Animals were divided into four groups of five each. On day 0, animals were anesthetized, fur of the dorsum of each animal was shaved and a wound area was created on the dorsal interscapular region 5 mm away from the ears using a circular colored rubber stamp. Full thickness skin from the demarcated area was excised including panniculus carnosus to get a wound area of approximately 400 mm². After wound creation experimental animals were randomly divided into four groups of five animals each. Except for Group I, all the groups received assigned treatment until complete closure of the wound had occurred. Body weights were monitored on days 0, 7, and 14. Wound area was measured on every alternative day of the study, i.e., on days 0, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, and 21. Percentage wound area was calculated. The study design is indicated in Table 1. Individual animal body weights were recorded on days 0, 7, and 14.

Table 1.

Experimental animal design for wound healing analysis in Wistar rats

Groups	Number of animals
Control	5
AM1122-100%	5
AM1122-75%	5
AM1122-50%	5
Total	20

Assessment of wound healing

The physical attributes of wound healing, namely, wound closure, were studied by tracing the raw wound. Wound area was measured by retracing the wound on every alternate day. The degree of wound healing was calculated:

% wound closure = \frac{\begin{matrix} Wound area on day 0 \\ - wound area on day N \end{matrix}}{Wound area on day 0} \times 100

where N is the corresponding day the wound area was measured.

Histopathological analysis

At the end of the study the respective groups wound tissue was collected and fixed in 10% formalin. After removing lipid debris by soaking in alcohol, the sections were dehydrated in increasing ethanol concentrations and the tissue samples were embedded in paraffin wax. The section which is 5–10 µm thick coronal section of wound tissue with a width of 2 mm was taken using a microtome. Slides were stained with hematoxylin and eosin (H&E). The stained tissue sections were observed under a motic digital microscope (40×) and the wound tissue region was photographed to assess the changes observed by different treatments.²¹

Statistical analysis

For determination of significant inter-group difference of each parameter, one-way analysis of variance (ANOVA) was carried out and Dunnet’s test was used for individual comparisons after significant ANOVA results. The differences with p < 0.05 were considered statistically significant. GraphPad Prism 8 software (GraphPad software, Inc., CA, USA) was used for statistical analysis.

Results and discussion

In the current study, A. indica and T. grandis extracts were successfully obtained through UAE, utilizing ethanol as the solvent. Ethanol’s polarity and solubility make it an effective choice for extracting bioactive compounds, while UAE provides a more advanced extraction technique known for its eco-friendly approach and efficiency in preserving the integrity of heat-sensitive phytochemicals. Comparing this with previous findings, UAE has been demonstrated to increase the extraction efficiency, particularly for plant secondary metabolites, while minimizing solvent usage and reducing extraction time. This is advantageous because, as other studies affirm, UAE allows for enhanced mass transfer, preserves sensitive compounds at lower temperatures, and minimizes degradation of bioactive components, an essential factor for wound-healing applications where the stability and activity of bio-compounds are critical.

Furthermore, UAE applies ultrasonic waves that stimulate plant cells gently, causing them to release active ingredients without compromising their structure. Prior research supports that UAE promotes the preservation and extraction of secondary metabolites such as terpenoids, saponins, alkaloids, and tannins, which are known to contribute significantly to wound healing due to their anti-inflammatory, antioxidant, and antibacterial properties. This study’s use of UAE aligns with the goals of sustainability and environmental protection by reducing reliance on organic solvents and improving the purity of the extracted compounds, a feature that enhances the suitability of A. indica and T. grandis extracts for biomedical applications²² (Table 2).

Table 2.

Phytochemical analysis of A. indica and T. grandis

Bioactive compounds	Response
Tannins	+
Saponins	–
Flavonoids	–
Cardiac glycosides	–
Terpenoids	+
Steroids	+
Alkaloids	+

Thin layer chromatography

The qualitative analysis using TLC in this study identified a variety of phytocompounds in the ethanolic, methanolic, and chloroform extracts of A. indica and T. grandis (Figure 1 and Table 3). The retention factor (Rf) values obtained from each solvent system indicate the presence of cardiac glycosides, flavonoids, saponins, steroids, tannins, and terpenoids. This approach of utilizing multiple solvent systems to extract and identify bioactive compounds is supported by recent studies that demonstrate TLC’s efficacy in profiling complex plant extracts.

Table 3.

TLC analysis of A. indica and T. grandis

Phytochemical constituents	Solvent system for TLC	Methanol extractRf (color spot)	Ethanol extractRf (color spot)	Chloroform extractRf (color spot)
Alkaloids	Ethyl acetate:methanol:water(81:11:8)	0.792(pink)	0.92(pink)	0.792(pink)
Flavonoids	Chloroform:ethanol:glacial acetic acid(94:5:1)	–	0.679(yellow)	–
Saponins	Chloroform:methanol:water(64:32:2)	–	–	–
Steroids	Chloroform:acetone(80:20)	0.685(green)	–	0.710(yellow)
Tannins	Chloroform:ethyl acetate:methanol(50:30:20)	0.9125(green)	–	0.875(yellow) 0.9(yellow)
Terpenoids	Toluene:ethyl acetate(93:7)	0.151(pink) 0.244(pink)	0.069(pink) 0.093(pink)	0.151 (pink) 0.255 (pink) 0.290 (pink) 0.476 (green) 0.588 (pink) 0.616 (pink)

For example, Khatoon et al.²³ utilized TLC to effectively separate and identify active compounds in medicinal plants, emphasizing the technique’s capability to detect flavonoids and terpenoids among other compounds. Their work also highlighted that different solvent systems optimize the separation of compounds, a method aligned with the current study’s use of varied mobile phases to enhance compound identification in A. indica and T. grandis.

Furthermore, Chen et al.²⁴ demonstrated that TLC is effective for preliminary screening of phytochemicals due to its cost-effectiveness, simplicity, and rapidity, especially when analyzing bioactive compounds with different polarities. They successfully used TLC to identify glycosides, tannins, and saponins in different plant extracts, emphasizing that such compounds can have antimicrobial and anti-inflammatory properties, which are beneficial for wound-healing applications. This supports the current study’s findings and suggests that the bioactive compounds identified may contribute to the therapeutic effects observed.

Bhale et al.²⁵ also highlighted that TLC is an excellent method for initial screening of plant-based compounds, noting that the technique is particularly suited to identifying and quantifying flavonoids and terpenoids, both of which play critical roles in wound healing due to their antioxidant and anti-inflammatory properties. Their study found that TLC could efficiently separate these bioactives, which have been linked to enhanced cellular repair and tissue regeneration in wounds.

In addition, the retention factor values observed in this study are consistent with those reported by Yadav et al.,²⁶ who studied the Rf values of various compounds in T. grandis. They reported similar Rf ranges for flavonoids and tannins, corroborating the identification methods used in the current research. Such alignment with recent literature strengthens the validity of the current study’s findings.

The identification of these bioactive compounds in A. indica and T. grandis suggests their potential as effective agents in wound healing. Secondary metabolites such as flavonoids and saponins have been shown to promote wound contraction and re-epithelialization, highlighting their importance in skin repair and regeneration.²⁷ This literature connection further suggests that the identified phytocompounds could provide valuable contributions to the fabric’s bioactivity in wound-care applications.

Gas chromatography profile of bio-consortia of A. indica and T. grandis

The bioactive compounds in the consortium extract of A. indica and T. grandis were separated and identified using GC-MS (Figure 2). The GC-MS analysis revealed the presence of eight compounds, as detailed in Table 4. Among these, silane was identified as a new compound that was not detected in the individual plant extracts but appeared in the combined consortium. Silane, a compound known for its application in fiber treatments, enhances the physical and mechanical properties of fibers significantly, particularly bamboo fibers. Previous studies have shown that silane treatment improves the adhesion and interfacial bonding between fibers, leading to superior tensile and flexural strength compared with untreated bamboo fabric.²⁸ The presence of silane in the extract is particularly beneficial for the fabrication process, as it improves the bonding of the bioactive compounds with the bamboo fabric, enhancing the overall properties of the fiber.

Table 4.

GC-MS analysis for molecular weight analysis of A. indica and T. grandis

S No.	Retention time	Molecule name	Molecular formula	Molecular weight (g/mol)
1.	1.229	2-Propanesulfinic acid, methyl ester	C₄H₁₀O₂S	122
2.	1.259	2-Propenenitrile, 2-chloro-	C₃H₂NCl	87
3.	1.289	4-Chlorobuten-3-yne	C₄H₃Cl	86
4.	1.329	Chloromethyl propanoate	C₄H₇O₂Cl	122
5.	1.429	Propanamide, 2-hydroxy-	C₃H₇O₂N	89
6.	1.509	Silane, dimethyl-	C₂H₈Si	60
7.	21.317	Silane, dimethyl (2-naphthoxy) octadecyloxy-	C₃₀H₅₀O₂Si	470
8.	22.217	Cholest-2-eno[2,3-B] indole, 4′-methoxy-	C₃₄H₅₁ON	489

Another notable compound identified in the extract is 2-propanesulfinic acid, which is known for its antioxidant properties. It plays a significant role in trapping free radicals, thus contributing to the overall antioxidant activity of the extract.²⁹ This compound supports the potential of the fabric as an antioxidant agent, which can aid in reducing oxidative stress in wounds and enhancing the healing process.

In addition, methyl esters, a byproduct of transesterification, were detected in the extract. Methyl esters are well known for their effective antibacterial and antifungal properties, which are valuable for wound-dressing applications. These properties can aid in preventing infection and promoting faster healing in treated wounds. The antibacterial and antifungal activities of methyl esters further contribute to the potential of the bamboo fabric treated with A. indica and T. grandis extracts for wound-care applications.

In summary, the identification of these compounds, particularly silane, 2-propanesulfinic acid, and methyl esters, highlights the multifunctional role of the bioactive extract in enhancing the mechanical, antioxidant, antibacterial, and antifungal properties of the bamboo fabric. These findings align with recent research on the synergistic effects of plant extracts and fiber treatments, suggesting that the fabric could be a promising candidate for wound-healing applications.³⁰

FTIR analysis of bioactives of A. indica and T. grandis

The functional group analysis by infrared spectrum was formed out in a fixed cell with an appropriate solvent. Since the UAE mixture was ethanolic extract, FTIR spectroscopy has been performed accordingly. The fixed cell size was about 0.1 mm thick and a spacer of 0.025 was used in the detection. The functional groups were analyzed using FTIR spectroscopy in the region of 4000–700 cm⁻¹ (Figure 3). FTIR characterization of A. indica and T. grandis exhibited possible aromatic rings/functional groups present in the bio-consortia. The absorbance peaks are absorbed at 11 different regions in the spectra are observed (Table 5). In the region between 4000 and 1800 cm^–1, the prominent band centered around 3295.81 cm^–1 designated to O–H stretching vibration of glycosidic structures. These O–H stretching vibrations could be overlapped by inter- and intra-hydrogen bonds that are available in polysaccharides. The bands around over 1800–1500 cm^–1 in turn designated to C=O stretching of fatty acids. In the region between 1350 and 1000 cm^–1 indicated C–N was assigned to amines and the bands around over 1450–1375 cm^–1 is assigned to –CH₃ as alkanes.³¹ In the region between 950 and 750 cm, two major bands C–X and C–H (aromatic) were assigned to chloride and aromatic proteins.

Table 5.

FTIR spectrum for A. indica and T. grandis

Peak frequency range (cm)	Functional group
3295.81	Alcohols, phenols (H-bonded) (O–H)
2920.25	Alkanes (C–H)
2851.06	Alkanes (C–H)
1791.78	Aldehyde (C=O)
1605.64	Alkene (C=C)
1378.45	Alkanes (C–H)–CH₃, S=O sulfones, sulfonyl chlorides, sulfates, sulfonamides
1316.68	Amines (C–N), C–X fluoride, S=O sulfones, sulfonyl chlorides, sulfates, sulfonamides
1239.22	Amines (C–N), C–X fluoride, S=O sulfones, sulfonyl chlorides, sulfates, sulfonamides
1148.66	Amines (C–N), C–X fluoride
1024.75	Amines (C–N), C–X fluoride

Antioxidant analysis of bio-consortia of A. indica and T. grandis extracts

Antioxidants are substances that can prevent or slow damage to cells caused by free radicals, unstable molecules that the body produces as a reaction to environmental and other pressures. In this study antioxidant activity was revealed by FRAP assay. The quantity of the hydroxyl groups at the aromatic ring binding site as well as the kind of substituent determine the antioxidant activity.³² With an observed color change from yellow to bluish green this estimates the ability of the antioxidants in the A. indica extracts to reduce Fe³⁺ to give Fe²⁺. Overall findings show that the extract can reduce Fe³⁺ ions in the ferricyanide complex are retrieved in polar solvents more readily than non-polar solvents.³³ Ascorbic acid was used as a reference point for comparing the outcomes. As tannins and saponins are present in T. grandis which results in its antioxidant property. This finding shows that T. grandis has a potent ability to combat illness brought on by free radicals.³⁴ It is proven that these plant extracts exhibit antioxidant property. Similarly, the consortium of A. indica and T. grandis also shows antioxidant activity (Figure 4).

Antibacterial activity assessment of dyed bamboo fabrics with A. indica and T. grandis extracts

The antibacterial properties of A. indica and T. grandis have garnered significant attention, with various studies investigating their effectiveness against both Gram-positive and Gram-negative bacteria (Figures 5 and 6 and Tables 6 and 7). In this study, the antibacterial activity of these plant extracts was tested on B. subtilis (Gram-positive) and E. coli (Gram-negative) using ethanolic extracts at concentrations of 50%, 75%, and 100%. The results indicated a progressive increase in the zone of inhibition, with the highest inhibition observed in the 100% extract samples (22.7 mm for B. subtilis and 22.5 mm for E. coli), confirming the potent antibacterial activity of A. indica and T. grandis.

Table 6.

Zone of inhibition measured for different concentration of A. indica and T. grandis against B. subtilis

Gram-positive bacteria: B. subtilis
Extract concentration (%)	Zone of incubation (diameter (mm))
50%	21
75%	22
100%	22.7

Table 7.

Zone of inhibition measured for different concentration of A. indica and T. grandis against E. coli

Gram-negative bacteria: E. coli
Extract concentration (%)	Zone of incubation (diameter (mm))
50%	21.2
75%	21.7
100%	22.5

These findings align with the general antibacterial activity of plant-based extracts, as observed in several other studies. For example, aloe vera and eucalyptus extracts exhibited significant antibacterial activity against both Gram-positive and Gram-negative bacteria, though the zones of inhibition were somewhat lower (18–21 mm) in comparison with the current study.³⁵ Similarly, extracts from Azadirachta indica (neem) were shown to inhibit bacterial growth effectively, with zones ranging from 15 to 23 mm depending on the bacterial strain and concentration.

The distinction between Gram-positive and Gram-negative bacteria, particularly in terms of cell wall structure, plays a crucial role in their response to antibacterial agents. Gram-positive bacteria possess a thicker peptidoglycan layer, making them generally more susceptible to certain plant extracts.³⁶ In contrast, Gram-negative bacteria have an additional outer membrane that can provide a layer of protection, often necessitating higher concentrations of active compounds for effective inhibition. The observed efficacy of A. indica and T. grandis against both types of bacteria suggests that these extracts could have broad-spectrum potential, a feature that is highly sought after in the development of natural antimicrobial agents.

The current study’s results, therefore, not only reinforce the antibacterial properties of these plants but also contribute to the growing body of evidence supporting the use of plant-based bioactives in combating bacterial infections. Moreover, future studies could explore the specific compounds responsible for this activity and their mechanism of action, as understanding the bioactive components can further optimize the development of natural antibacterial agents for therapeutic applications.

Computer color matching system

The untreated fabric and consortium treated fabrics were characterized by spectral reflectance. The color strength of a dyed fabric is measured using a spectrophotometer. Absorption coefficient (K) and scattering coefficient (S) and their ratio (K/S) is a measure of color strength. The untreated dyed sample is compared with a treated sample.³⁷ It has been analyzed that the K/S value of undyed bamboo fabric is 13 and the K/S value of bamboo fabric coated with bio-consortia extract is 15, which increases with the rise in concentration of the extracted solution. It indicates that shade depth depends on the concentration of the bio-consortia-treated samples. However, after 75% concentration, the K/S value of A. indica and T. grandis levels off, and that was why no further increase in K/S value was observed for sample treated with 100% A. indica and T. grandis.

Fragrance intensity testing

A fragrance intensity test was performed using a subjective procedure where the fragrance intensity was rated on a Likert scale from 0 (no aroma) to 7 (high aroma intensity) (Figure 7). The results revealed that even without washing, the fragrance intensity gradually declined over time in natural environmental conditions. After 4 weeks, the fragrance intensity was rated 4 on the scale, suggesting moderate fragrance retention. Notably, fabric treated with 100% extract exhibited higher fragrance ratings compared with the 75% and 50% concentrations, likely due to better binding properties between the bioactive compounds and the fabric fibers, supporting the premise of improved fragrance retention with higher extract concentration.

Regarding the longevity of fragrance retention, the fabric maintained its fragrance for up to 20 washes and continued to exhibit detectable fragrance for more than 4 weeks under natural conditions. However, the specific chemical mechanism through which the bioactive compounds preserve the fragrance remains unclear. Further studies should focus on investigating the molecular interactions between the bioactive compounds and the fabric fibers, which could offer deeper insights into the durability and degradation mechanisms of the fragrance over time.

Similar findings in the literature suggest that microencapsulation techniques, such as complex coacervation, play a crucial role in enhancing fragrance retention in textiles, improving the washing durability of treated fabrics. These encapsulation methods also provide controlled release mechanisms that contribute to sustained fragrance release under real-world conditions, thereby promoting the development of long-lasting, hygienic, and functional textile products.

Olfaction China GB test

The admissible smell of A. indica and T. grandis treated fabric was determined through the olfaction method (Table 8). As a result of the analysis, it is observed that our fabric satisfies GB/T 18885-2009 (Technical specification of ecological textiles code) and GB/T 20097-2006 (Protective clothing code) standards (ChineseStandard.net 2010). It is observed that the odor of treated fabric is admissible and aromatic with a good odor.³⁸

Table 8.

Olfaction China GB test analysis of treated fabric

Tested material	Type of smell	Herbal smell	Aromatic smell	Good/bad odor
Undyed fabric	No fragrance	No	No	No
50% dyed fabric	Peculiar	Yes	Yes	Good
75% dyed fabric	Peculiar	Yes	Yes	Good
100% dyed fabric	Peculiar	Yes	Yes	Strong

Wound closure in Wistar rats treated with A. indica and T. grandis wound-dressing material

Effect of body weight

The body weight results were monitored every 7 days in the study with results as listed in Table 9. During day –1, body weight of all the groups with different treatment animals were observed and recorded for all the groups. From day 7 to day 14 there is no significant difference in the body weight was observed and recorded (Figure 8).

Table 9.

Effect of bio extract formulation of A. indica and T. grandis on body weight of treated and untreated Wistar rats (g)

Groups	Treatment	Body weight (g)
Groups	Treatment	Day 0	Day 1	Day 7	Day 14
I	Control	195 ± 0.21	198 ± 0.18	209 ± 0.87	217 ± 0.84
II	AM1122-100%	196 ± 0.32	197 ± 0.54	210 ± 0.75	216 ± 0.74
III	AM1122-75%	195 ± 0.39	198 ± 0.56	209 ± 0.54	218 ± 0.84
IV	AM1122-50%	194 ± 0.43	197 ± 0.65	210 ± 0.86	217 ± 0.76

Values are mean ± standard deviation, n = 5.

p < 0.05 versus Group I (control), ANOVA followed by Dunnett’s multiple comparison test.

Assessment of wound healing

The wound-healing results were recorded on alternate days throughout the study as indicated by Table 10 and Figure 9. During days 5–21, significance variation in wound area in bio extract formulation of AIT1122 at three different concentrations 100%, 75%, and 50% were observed and recorded (Figure 10), whereas bio extract formulation treated with A. indica and T. grandis (AIT1122) groups showed a more significant difference when compared with the control group with regards to percentage of wound closure^39,40 (Table 11 and Figures 10 and 11)

Table 10.

Effect of bio extract formulation of A. indica and T. grandis on the wound area of treated and untreated Wistar rats

Days	Wound area (mm²)
Days	Group I(Normal control)	Group II(AM1122-100%)	Group III(AM1122-75%)	Group IV(AM1122-50%)
0	400 ± 0	400 ± 0	400 ± 0	400 ± 0
1	374 ± 2.92	373.2 ± 5.4	374.8 ± 3.35	373.8 ± 2.17
3	288.6 ± 1.14	279.8 ± 1.64	283.6 ± 2.7	293.6 ± 1.67
5	265.6 ± 2.07	263.6 ± 3.05	263.6 ± 1.82	272.6 ± 2.07
7	225.4 ± 2.41	233.4 ± 3.21	238 ± 2.24	255.2 ± 3.11
9	172.6 ± 2.7	162.8 ± 1.3	172.4 ± 1.67	184 ± 2.45
11	138.2 ± 1.48	123.4 ± 2.79	136 ± 2.12	145 ± 3.39
13	77.2 ± 2.17	65.8 ± 2.28	75.4 ± 3.36	83.6 ±2.41
15	53.2 ± 2.17	45.6 ± 2.07	52.6 ± 3.65	64.2 ± 2.86
17	27.2 ± 1.92	23.6 ± 2.88	35.2 ± 1.48	44.4 ±2.7
19	11.4 ± 1.14	8 ± 1	15.6 ± 3.44	24.6 ± 1.95
21	2.6 ± 0.89	1.4 ± 0.55	6.4 ± 1.14	10.8 ± 3.27

Values are mean ± standard deviation, n = 5.

p < 0.05 versus Group I (control), ANOVA followed by Dunnett’s multiple comparison test.

Table 11.

Wound area closure analysis of Wistar rats on treatment with A. indica and T. grandis bio extract

Days	Wound closure (mm²)
Days	Group I(Normal control)	Group II(AM1122-100%)	Group III(AM122-75%)	Group IV(AM1122-50%)
0	0 ± 0	0 ± 0	0 ± 0	0 ± 0
1	6.5 ± 0.73	6.7 ± 1.35	6.3 ± 0.84	6.55 ± 0.54
3	27.85 ± 0.29	30.05 ± 0.41	29.1 ± 0.68	26.6 ± 0.42
5	33.6 ± 0.52	34.1 ± 0.76	34.1 ±0.45	31.85 ± 0.52
7	43.65 ± 0.6	41.65 ± 0.8	40.5 ± 0.56	36.2 ±0.78
9	56.85 ± 0.68	59.3 ± 0.33	56.9 ± 0.42	54 ± 0.61
11	65.45 ± 0.37	69.15 ± 0.7	66 ± 0.53	63.75 ± 0.85
13	80.7 ± 0.54	83.55 ± 0.57	81.15 ± 0.84	79.1 ± 0.6
15	86.7 ± 0.54	88.6 ± 0.52	86.85 ± 0.91	83.95 ± 0.72
17	93.2 ± 0.48	94.1 ± 0.72	91.2 ± 0.37	88.9 ± 0.68
19	97.15 ± 0.29	98 ± 0.25	96.1 ± 0.86	93.85 ± 0.49
21	99.35 ± 0.22	99.65 ± 0.14	98.4 ± 0.29	97.3 ± 0.82

Values are mean ± standard deviation, n = 5.

p < 0.05 versus Group I (control), ANOVA followed by Dunnett’s multiple comparison test.

Histopathological study

The histopathological features of the tissue of all group animals were examined under motic microscope (10×). From (Figure 12). Group I (Control) animals showed inflammatory cells, reduced collagen fibers, and blood vessels; there is also the presence of visible scar tissue. Groups II, III, and IV showed a cell regeneration of the tissue which was evident by increased collagen fibers and blood vessels; it showed a higher significance when compared with the control Group I.

Conclusion

In conclusion, our study has provided compelling evidence for the potential of bamboo fabric treated with A. indica and T. grandis as a multifaceted material for wound care, antibacterial protection, and fragrance retention. Our key findings include the significant antibacterial activity of the bio-extract-treated fabrics against both E. coli and B. subtilis, which suggests the fabric’s ability to inhibit microbial growth and prevent bacterial adhesion. In addition, we observed that the fabric retained a pleasant fragrance over time, attributed to the enhanced binding properties of the bioactive compounds with the bamboo fibers. One of the innovative aspects of this study is the integration of plant-based bioactive compounds, specifically from A. indica and T. grandis, into textile materials, which offers new possibilities for the development of functional and sustainable fabrics in the healthcare industry. This approach not only enhances the antibacterial properties of the fabric but also contributes to prolonged fragrance retention, an added benefit for consumer hygiene products.

Our hypothesis, that the treated fabric would exhibit antibacterial properties and enhanced wound-healing capabilities, was supported by our findings, including the significant improvement in wound closure observed in preclinical tests with Wistar rats. This suggests that the bio-extract-treated fabric could play a crucial role in wound management.

Looking forward, we envision future work focused on scaling up these treatments for industrial applications. Further research is necessary to explore the molecular interactions between the bioactive compounds and the fabric fibers to optimize both the fragrance retention and antibacterial effectiveness. In addition, long-term stability testing under real-world conditions will be crucial to ensure that these treated fabrics maintain their functional properties over time, particularly after repeated washing or extended use.

In this study, the findings on the antibacterial properties and fragrance retention of bamboo fabrics treated with A. indica and T. grandis can be more comprehensively understood by directly comparing them with those of other researchers in the field. For example, while our results demonstrated significant antibacterial activity against E. coli and B. subtilis, it is essential to compare these outcomes with similar studies using different plant-based extracts or textiles. Previous studies have shown that fabrics treated with plant extracts such as aloe vera and Calendula officinalis also exhibit antimicrobial properties.²⁷ However, the differences in their antibacterial efficacy could be attributed to the specific bioactive compounds, extraction methods, and fabric types used in each study.

Furthermore, the fragrance retention observed in our fabric can be compared with other research that has examined similar applications. For instance, fabrics treated with essential oils or other plant extracts (such as those in the work by Wang et al.²⁹) also retained fragrance, but the duration of effectiveness and the impact of repeated washing may differ due to the varying binding mechanisms of the active compounds. Our study shows that A. indica and T. grandis extracts exhibit better binding properties, which may offer advantages in long-term fragrance retention compared to other studies.

To improve the scientific rigor of our work, it would be beneficial to perform a direct comparison of the forecasting results of these findings with those of other works. This would help in determining the relative performance of our treated fabrics against established benchmarks, offering insights into the most effective plant extracts for wound care and textile treatments. Future research should include a broader comparative analysis to identify potential areas for further optimization and refinement of bioactive textile applications.

This study paves the way for further exploration of plant-based treatments in textiles, and it could lead to the development of innovative, eco-friendly materials for various healthcare applications, contributing to the advancement of the textile industry’s sustainability efforts

Footnotes

Acknowledgment

This research paper is contributed to by the first three authors pursuing B. Tech Biotechnology in their final years. The authors gratefully acknowledge the Department of Biotechnology and Sri Shakthi Institute of Engineering and Technology for providing an ambient environment for the successful completion of the project.

Declaration of conflicting interests

The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Funding

The author(s) received no financial support for the research, authorship, and/or publication of this article.

ORCID iD

J Bindhu

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