Preparation of Cellulose Scaffolds from Leaves for Wound Regeneration
Surendra Pratap Chakravarti
Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Anil Kumar Gangwar
Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Sangeeta Devi Khangembam *
Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Vipin Kumar Yadav
Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Yogendra Singh
Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Rajesh Kumar Verma
Department of Veterinary Clinical Complex, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Manoj Kumar Verma
Department of Animal Genetics and Breeding, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Rabindra Kumar
Department of Veterinary Gynaecology and Obstetrics, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Prafull Kumar Singh
Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Anil Singh
Department of Veterinary Clinical Complex, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
Raj Kapoor Verma *
Department of Livestock Production Management, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Ayodhya (UP), India.
*Author to whom correspondence should be addressed.
Abstract
Plant-derived tissue decellularisation offers a potentially accessible route for producing cellulose-rich scaffolds for regenerative research. This study prepared leaf-based scaffolds through sequential surface decutinisation and chemical decellularisation. Fresh leaves were treated with an n-hexane-diethyl ether mixture, followed by 4% sodium dodecyl sulphate for 120 h, 2% Triton X-100 for 48 h, and 4% sodium hypochlorite for 12 h. The processed leaves were washed repeatedly with sterile deionised water. Scaffold decellularisation and structural preservation were assessed by gross examination, haematoxylin and eosin staining, DAPI fluorescence staining, scanning electron microscopy, DNA quantification, and an in vitro haemocompatibility assay. Native and decellularised dry leaf tissues contained 105.36 ± 1.03 ng/mg and 35.27 ± 0.52 ng/mg total DNA, respectively. Histological and DAPI observations indicated substantial removal of cellular and nuclear material, while scanning electron microscopy showed preservation of the general porous architecture and stomatal features. Haemolysis was significantly lower for decellularised leaf tissue than for native leaf tissue, with values of 1.90 ± 0.53% and 30.71 ± 3.67%, respectively. These findings indicate that sequential treatment with sodium dodecyl sulphate and Triton X-100 can produce cellulose-rich leaf scaffolds with reduced residual DNA and preliminary blood compatibility. Further mechanical, cytocompatibility, degradation, antimicrobial, and in vivo wound-healing evaluations are required before practical regenerative use can be considered.
Keywords: Plant-derived scaffold, leaf decellularisation, cellulose matrix, wound regeneration, tissue engineering, sodium dodecyl sulphate, Triton X-100, DAPI staining, scanning electron microscopy, haemocompatibility