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Characterization of Glutaraldehyde Crosslinked Decellularized Caprine Gall Bladder Scaffolds Prepared Using Sapindus mukorossi Fruit Pericarp Extract

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Abstract

Purpose

The commonly used decellularization agent such as sodium dodecyl sulfate, Triton X-100, and sodium deoxycholate has the potential to cause residual cytotoxic effects in the extracellular matrix. The present study was conducted to establish a protocol for decellularization of caprine gall bladder using aqueous soap nut (Sapindus mukorossi) pericarp extract.

Methods

The gall bladder tissues were decellularized using 5% aqueous extract of Sapindus mukorossi fruit pericarp under continuous agitation on the magnetic stirrer and crosslinked with glutaraldehyde. The efficacy of the decellularization process was verified with the help of histology, scanning electron microscopy, DAPI staining, and DNA content estimation. The physico-chemical properties of crosslinked scaffolds were evaluated by enzymatic and non-enzymatic degradation, free protein, free amino group, fixation index, hydroxyproline, and moisture content.

Results

The gall bladder tissue was completely decellularized at 72-h interval. The weight loss of decellularized glutaraldehyde crosslinked scaffolds following non-enzymatic, and collagenase digestion significantly increased with the digestion time interval. The free protein and free amino acid group concentration of acellular gall bladder matrix was significantly (P < 0.05) higher than crosslinked tissues collected at different crosslinking time intervals. The fixation index was highest in tissues crosslinked for 72 h. The moisture content at the 72 h crosslinking interval was significantly lower (P < 0.05). The free hydroxyproline content in the crosslinked acellular matrices was nil and did not show any band pattern in SDS-PAGE gel.

Conclusion

The findings from this study indicate that soap nut pericarp extract is a viable option for decellularizing caprine gall bladder without any toxicity.

Lay Summary and Future Work

The commonly used decellularization agent such as sodium dodecyl sulfate, Triton X-100, and sodium deoxycholate has the potential to cause residual cytotoxic effects in the extracellular matrix. So, we used aqueous soap nut (Sapindus mukorossi) pericarp extract for decellularization of caprine gall bladder. The efficacy of the decellularization process was verified with the help of histology, scanning electron microscopy, DAPI staining, and DNA content estimation. The physico-chemical properties were evaluated by enzymatic and non-enzymatic degradation, free protein, free amino group, fixation index, hydroxyproline, and moisture content. The gall bladder tissue was completely decellularized at 72-h interval. The findings from this study indicate that Sapindus mukorossi fruit pericarp extract is a viable option for decellularizing caprine gall bladder without any toxicity.

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Acknowledgements

The authors are thankful to their respective universities and institute for the facilities provided during the course of study.

Funding

Hon’ble Vice chancellor of the University provided all necessary facilities to conduct this study.

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Authors and Affiliations

  1. Department of Veterinary Surgery and Radiology, College of Veterinary Science and Animal Husbandry, Acharya Narendra Deva University of Agriculture and Technology, Kumarganj, Ayodhya, 224 229, Uttar Pradesh, India

    Amit Kumar Sachan, Anil Kumar Gangwar & Sangeeta Devi Khangembam

  2. Division of Surgery, I.V.R.I., Izatnagar, Uttar Pradesh, India

    Naveen Kumar

Authors
  1. Amit Kumar Sachan
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  2. Anil Kumar Gangwar
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  3. Sangeeta Devi Khangembam
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  4. Naveen Kumar
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Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by AKS, AKG, and SDK. Histological study was facilitated by NK. The first draft of the manuscript was written by AKG. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Anil Kumar Gangwar.

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Sachan, A.K., Gangwar, A.K., Khangembam, S.D. et al. Characterization of Glutaraldehyde Crosslinked Decellularized Caprine Gall Bladder Scaffolds Prepared Using Sapindus mukorossi Fruit Pericarp Extract. Regen. Eng. Transl. Med. 9, 240–248 (2023). https://doi.org/10.1007/s40883-022-00276-9

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  • DOI: https://doi.org/10.1007/s40883-022-00276-9

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