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Autologous Culture Expanded Iliac Crest Chondrocytes in Chitosan Hyaluronic Acid Dialdehyde Gel Regenerate Caprine Growth Plate

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Abstract

Purpose

Allogenic and autologous chondrocytes have been used to reconstitute damaged growth plates in small and large animals with variable success. We evaluated the efficacy of growth plate chondrocytes in a chitosan-hyaluronic acid dialdehyde (CHDA) hydrogel in repairing damaged growth plates using an immature goat model (n = 4).

Methods

Chondrocytes were harvested from the ipsilateral iliac crest cartilage and expanded in vitro. The culture-expanded cells were seeded in a CHDA hydrogel prior to transplantation. A physeal defect measuring approximately 1 cm3 was created in the proximal medial tibial physis, and cell-seeded hydrogel was transplanted into the defect. One animal that received green fluorescent protein-labeled cells (to determine the fate of transplanted cells) was sacrificed at 2 months, while the others were sacrificed at 6 months. The outcome was assessed by histology and radiographs.

Results

Our results indicated the presence of transplanted cells in the short term. The histology of the transplanted growth plate was comparable to normal and demonstrated a columnar arrangement with endochondral ossification. The mean tibial length of the transplanted limb was 18.63 (16.5 to 20.3), while that of the contralateral limb was 18.58 (16.5 to 20.4). Mean tibial valgus at the transplanted and contralateral sides were 6.56° (5.2 to 8.9) and 4.63° (1.4 to 7.4), respectively, and significantly different when compared to the defect alone, which had a varus deformity of 2.9° (0.2 to 5.6).

Conclusion

This study demonstrates the feasibility of encapsulating autologous chondrocytes in a novel hydrogel for growth plate regeneration, thereby preventing bony bridge formation and varus deformity.

Layman Summary

In children, growth plate cartilage is responsible for the growth of long bones. The structure is fragile, such that any injury or infection can interfere with normal growth, leading to inferior quality bone quality. If not treated early, it often leads to permanent disability; multiple corrective surgeries and time-consuming lengthening procedures are required to correct the deformity. In this study, using a young goat model, we tested if transplanting autologous growth plate cells at the defect site can restore normal bone growth, eliminating the need for laborious surgical procedures. Our results showed that it is likely to establish normal growth of the bone without bone abnormality using this technique.

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Data Availability

All data generated or analyzed during this study are included in this article.

Materials Availability

All data generated or analyzed during this study are included in this article.

Code availability

Not applicable.

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Acknowledgements

The authors would like to thank Dr. Michael P. Marino and Prof. Jakob Resier from Louisiana State University Health Sciences Center, New Orleans, LA, USA, for their technical assistance in the production of lentiviral vectors. The authors would also like to thank the technicians from the department of radiology and histopathology (CSCR) and central animal facility at Christian Medical College, Vellore. In addition, the authors acknowledge Dr. Sundarrajan, Ms. Legasri, Ms. Janani, Dr. Sangeet Gangadharan, and Dr. Vishak Manoj for their assistance during the animal surgeries.

Funding

This study was funded under the Indo-Danish call (BT/IN/DENMARK/02/PDN/2011 DATED26/05/11), Department of Biotechnology, Government of India.

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Authors and Affiliations

  1. Center for Stem Cell Research, a Unit of InStem Bengaluru, Christian Medical College, Vellore, 632002, Tamil Nadu, India

    Vrisha Madhuri, Sowmya Ramesh, Karthikeyan Rajagopal, Sanjay K. Chilbule & Noel Malcolm Walter

  2. Paediatric Orthopedics Unit, Christian Medical College, Vellore, 632004, Tamil Nadu, India

    Vrisha Madhuri, Sowmya Ramesh, Karthikeyan Rajagopal & Sanjay K. Chilbule

  3. Sree Chitra Tirunal Institute of Medical Sciences and Technology, Trivandrum, Kerala, India

    Prabha. D. Nair

  4. Department of Forensic Medicine, Christian Medical College, Vellore, 632 004, Tamil Nadu, India

    Noel Malcolm Walter

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  1. Vrisha Madhuri
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  2. Sowmya Ramesh
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  3. Karthikeyan Rajagopal
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Contributions

VM performed all the animal surgeries, measurements, and histological bits; SR and KR performed all in vitro experiments and data analysis and assisted during animal surgeries; SKC assisted during animal surgeries; PDN provided the scaffold; NMW performed all histological interpretations.

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Correspondence to Vrisha Madhuri.

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The necessary IRB and animal ethics approval were obtained prior to the study. This article consists of data involving large animal.

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Madhuri, V., Ramesh, S., Rajagopal, K. et al. Autologous Culture Expanded Iliac Crest Chondrocytes in Chitosan Hyaluronic Acid Dialdehyde Gel Regenerate Caprine Growth Plate. Regen. Eng. Transl. Med. 9, 397–406 (2023). https://doi.org/10.1007/s40883-022-00289-4

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