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 Table of Contents  
DPU: INTERDISCIPLINARY CONFERENCE
Year : 2020  |  Volume : 7  |  Issue : 5  |  Page : 95-97

Chitosan and stemcells: A synchrony for regeneration


1 Departments of Oral and Maxillofacial Surgery, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
2 Department of Pharmaceutical Chemistry, Dr. D. Y. Patil Institute of Pharmaceutical Sciences and Research, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India
3 Department of Research, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune, Maharashtra, India

Date of Web Publication26-Feb-2020

Correspondence Address:
Lakshmi Shetty
Department of Oral and Maxillofacial Surgery, Dr. D. Y. Patil Dental College and Hospital, Dr. D. Y. Patil Vidyapeeth, Pimpri, Pune - 411 018, Maharashtra
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/jdrr.jdrr_75_19

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  Abstract 


Background: Stem cells have infinite self-renewing capacity and potential for differentiation and regeneration. Human dental pulp stem cells (hDPSCs) from the extracted tooth are a rich source of mesenchymal stem cells, which can regenerate into osteogenic, chondrogenic, or adipogenic lineages. Chitosan which is a porous structure may have the ability to house these stem cells for regeneration. This study was to determine the effectiveness of growth of hDPSCs in the chitosan scaffolds. Methodology: The study was conducted in the Regenerative Medicine Laboratory, Dr. D.Y. Patil Dental College and Hospital in coordination with Department of Oral and Maxillofacial Surgery, Dr. D.Y. Patil Dental College and Hospital, and Dr. D.Y. Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune. The chitosan scaffolds were prepared at Dr. D.Y. Patil Institute of Pharmaceutical Sciences and Research. Twenty extracted human dental pulp were used for stem cells differentiation. Results: The chitosan scaffold showed an excellent growth of chondrogenic cells per unit in × 40 magnification, and hence, this sets a benchmark for hDPSC study in India. Conclusion: hDPSCs in porous chitosan scaffolds help us to regenerate chondrogenic cells and will initiate the research in an interdisciplinary approach toward cartilage regeneration.

Keywords: Chitosan, regeneration, scaffolds


How to cite this article:
Shetty L, Badhe R V, Bhonde R, Waknis P, Londhe U. Chitosan and stemcells: A synchrony for regeneration. J Dent Res Rev 2020;7, Suppl S2:95-7

How to cite this URL:
Shetty L, Badhe R V, Bhonde R, Waknis P, Londhe U. Chitosan and stemcells: A synchrony for regeneration. J Dent Res Rev [serial online] 2020 [cited 2020 Apr 2];7, Suppl S2:95-7. Available from: http://www.jdrr.org/text.asp?2020/7/5/95/278911

Editor: Dr. Pradnya Kakodkar





  Introduction Top


Chitosan is a polysaccharide of significant importance in the field of regenerative medicine. It is a polymer with multifunctional properties and enormous use.[1] Chitosan is biocompatible, and it is biodiversified applications to make it ideal in the regeneration of hard-tissue structures.[2] Mesenchymal stem cells have the ability to differentiate to osteogenic, chondrogenic, adipogenic, and neurogenic lineages.[3] In the field of oral and maxillofacial surgery, temperomandibular disorders seem to be the area of research where extensive treatment modalities both clinical, surgical, and research have been time tested, and stem cells seem to be the answer for it. Human dental pulp stem cells (hDPSCs) seem to be a platform for chondrogenic cells. This study was the need of the hour to study the in vitro growth of hDPSCs in the chitosan scaffolds to see the amount of chondrogenic cells generated to regenerate cartilage cells.


  Methodology Top


The study was conducted in the Regenerative Medicine Laboratory, Dr. D.Y. Patil Dental College and Hospital in coordination with Department of Oral and Maxillofacial Surgery, Dr. D.Y. Patil Dental College and Hospital, and Dr. D.Y. Patil Institute of Pharmaceutical Sciences and Research, Pimpri, Pune.

Study sample

Twenty samples of asymptomatic orthodontic extracted premolar teeth were taken after written informed Institutional Committee stem cell research approved the consent duly signed by the participants.

The human dental pulp was extirpated from the above samples under sterile conditions [Figure 1].
Figure 1: Pulp

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  • Phase 1: Basic research


    1. Extraction of teeth and removal of pulp
    2. In vitro cultivation of hDPSCs.


  • Phase 2: Preparation of chitosan scaffold
  • Phase 3: Growth of hDPSCs on chitosan scaffold and determination of chondrogenic differentiation.


Chitosan scaffold preparation

Chitosan Scaffold was done by the method of freeze-drying. Briefly, chitosan was dissolved in 0.2 M glacial acetic acid, and the solution was stored for 1 day at room temperature. Twenty-four hour later, the solution was poured into stainless steel mold and stored in deep freezer at −70°C next 5 days. The first phase was freezing phase, and the second pwwhase was warm-up vacuum pump phase, and then, chitosan scaffold was prepared.[4]


  Results Top


The hDPSCs derived from the study sample differentiated into colony-forming units of cluster of hDPSCs with chondrogenic differentiation for 21 days. The chitosan scaffold showed an excellent growth of chondrogenic cells per unit in × 40 magnification, and hence, this sets a benchmark for hDPSC study in India. Chondrogenic differentiation was determined by Safranin O Stain [Figure 2].
Figure 2: Cartilage

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  Discussion Top


Chitosan particles have the capacity of three-dimensional (3D) hard-tissue regeneration. In the field of oral and maxillofacial surgery, regeneration of bone and cartilage holds a lot of importance. Chitosan can be manufactured in a porous structure to allow for cell seeding. This space created by the porous structure allows for cell proliferation, migration, and the exchange of nutrients. The porosity of these scaffolds is beneficial for angiogenesis and function of the tissue regeneration.[5],[6] There is least foreign body reactionin vivo and specific reactions.[7] Chitosan used for the regeneration is ideal for the treatment of skeletal and joint diseases.[8] Chondrogenesis is the process where chondrogenic cells proliferate from mesenchymal cells and chondrocytes; at the end of the bone defect, these cells differentiate into mature osteocytes. Mesenchymal stem cells from dental pulp tissues can be cryopreserved, used whenever needed for regenerative therapeutics.[9] The disorders which are cured by hDPSCs are diabetes, neurological diseases, immunodeficiency diseases, and diseases of bone and cartilages.[10],[11] This study has made us aware that the regeneration of cartilage like cells would add on to our future research in temperomandibular joint cartilage regeneration.

Chitosan/nanohydroxyapatite composites have been more relevant for tissue engineering, because of its ability to induce a good proliferative response in osteoblasts, and in a tibial defect in a rabbit, it showed a good bone regeneration at 8 weeks seen by microcomputerized tomography.[12]

The defects in the joint, due to rheumatoid arthritis and dehydration of cartilage tissue in the entire body or by the lifting of heavy loads with the corresponding joint wear, lead to the total dependence of joint replacement therapy. This particular stem cell therapeutics of our study would eradicate the surgical option for a prosthesis in temporomandibular joint.

Collagen type II, protein in cartilage tissue, which enhances the adhesion and formation of chondrocytes in vitro regeneration.[13] Collagen II and chondroitin sulfate induce stem cells to differentiate to chondrocytes in vitro,[14] and similar results have been observed in our study too. To follow the seeding cells on chitosan composites, the next step in regeneration is the formation of functional tissue; collagen II expression by chondroblasts and chondrocytes is a determinant factor in cartilage formation, and it has been observed in the glycerophosphate-chitosan hydrogel with silk fibrils, where chondrocyte phenotype is maintained for the expression of glycosaminoglycans and type II collagen in vitro, as obtained with alginate and fibroin in chitosan hydrogels.[15] Extracellular matrix deposition is a key factor to recognize biocompatibility and normal cell function, in addition a 3D matrix for growth tissue by proliferation and differentiation of precursor cells. In the case of chondroblast and chondrocytes, production of collagen II is a functional tissue determinant, and this has been found in glycerophosphate-chitosan hydrogel and silk fibrils, which stimulate the production of collagen II and glycosaminoglycans by chondrocytes in vitro.[16]

The polylactide acid-chitosan with collagen provides a laminate matrix with mechanical properties similar to cartilage, but in addition, they have worked as a support for chondrocytes from rabbit cartilage,[17] and this opens the possibility to use chitosan composites in the regeneration of cartilage defects as in the case of arthritis or joint cartilage damage from aging.[18]


  Conclusion Top


This study concludes and allows for understanding the potential properties of chitosan composites in hard-tissue regeneration. In summary, chitosan and stem cells provide physical, chemical mechanical support, and differentiation, with the corresponding biocompatibility to induce the cartilage tissue regeneration.

Acknowledgments

I would like to thank Mr. Avinash Kharat, scientist from the regenerative medicine laboratory for his guidance and help.

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Kumar MN, Muzzarelli RA, Muzzarelli C, Sashiwa H, Domb AJ. Chitosan chemistry and pharmaceutical perspectives. Chem Rev 2004;104:6017-84.  Back to cited text no. 1
    
2.
Muzzarelli RA, Guerrieri M, Goteri G, Muzzarelli C, Armeni T, Ghiselli R, et al. The biocompatibility of dibutyryl chitin in the context of wound dressings. Biomaterials 2005;26:5844-54.  Back to cited text no. 2
    
3.
Potdar PD, Jethmalani YD. Human dental pulp stem cells: Applications in future regenerative medicine. World J Stem Cells 2015;7:839-51.  Back to cited text no. 3
    
4.
Garg T, Chanana A, Joshi K. Preparation of chitosan scaffolds for tissue engineering. IOSR Jr Pharmac 2012;2:72-3.  Back to cited text no. 4
    
5.
Madihally SV, Matthew HW. Porous chitosan scaffolds for tissue engineering. Biomaterials 1999;20:1133-42.  Back to cited text no. 5
    
6.
Ko YG, Kawazoe N, Tateishi T, Chen G. Preparation of chitosan scaffolds with a hierarchical porous structure. J Biomed Mater Res B Appl Biomater 2010;93:341-50.  Back to cited text no. 6
    
7.
VandeVord PJ, Matthew HW, DeSilva SP, Mayton L, Wu B, Wooley PH. Evaluation of the biocompatibility of a chitosan scaffold in mice. J Biomed Mater Res 2002;59:585-90.  Back to cited text no. 7
    
8.
Neves SC, Moreira Teixeira LS, Moroni L, Reis RL, Van Blitterswijk CA, Alves NM, et al. Chitosan/poly (epsilon-caprolactone) blend scaffolds for cartilage repair. Biomaterials 2011;32:1068-79.  Back to cited text no. 8
    
9.
Huang AH, Snyder BR, Cheng PH, Chan AW. Putative dental pulp-derived stem/stromal cells promote proliferation and differentiation of endogenous neural cells in the hippocampus of mice. Stem Cells 2008;26:2654-63.  Back to cited text no. 9
    
10.
Davila JC, Cezar GG, Thiede M, Strom S, Miki T, Trosko J. Use and application of stem cells in toxicology. Toxicol Sci 2004;79:214-23.  Back to cited text no. 10
    
11.
Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal Human Dental Pulp Stem Cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A 2000;97:13625-30.  Back to cited text no. 11
    
12.
Lee JS, Baek SD, Venkatesan J, Bhatnagar I, Chang HK, Kim HT, et al.In vivo study of chitosan-natural nano hydroxyapatite scaffolds for bone tissue regeneration. Int J Biol Macromol 2014;67:360-6.  Back to cited text no. 12
    
13.
Choi B, Kim S, Lin B, Li K, Bezouglaia O, Kim J, et al. Visible-light-initiated hydrogels preserving cartilage extracellular signaling for inducing chondrogenesis of mesenchymal stem cells. Acta Biomater 2015;12:30-41.  Back to cited text no. 13
    
14.
Choi B, Kim S, Lin B, Wu BM, Lee M. Cartilaginous extracellular matrix-modified chitosan hydrogels for cartilage tissue engineering. ACS Appl Mater Interfaces 2014;6:20110-21.  Back to cited text no. 14
    
15.
Sheehy EJ, Mesallati T, Vinardell T, Kelly DJ. Engineering cartilage or endochondral bone: A comparison of different naturally derived hydrogels. Acta Biomater 2015;13:245-53.  Back to cited text no. 15
    
16.
Mirahmadi F, Tafazzoli-Shadpour M, Shokrgozar MA, Bonakdar S. Enhanced mechanical properties of thermosensitive chitosan hydrogel by silk fibers for cartilage tissue engineering. Mater Sci Eng C Mater Biol Appl 2013;33:4786-94.  Back to cited text no. 16
    
17.
Yin D, Wu H, Liu C, Zhang J, Zhou T, Wu J, et al. Fabrication of composition-graded collagen/chitosan–polylactide scaffolds with gradient architecture and properties. React Funct Polym 2014;83:98-106.  Back to cited text no. 17
    
18.
Hao T, Wen N, Cao JK, Wang HB, Lü SH, Liu T, et al. The support of matrix accumulation and the promotion of sheep articular cartilage defects repairin vivo by chitosan hydrogels. Osteoarthritis Cartilage 2010;18:257-65.  Back to cited text no. 18
    


    Figures

  [Figure 1], [Figure 2]



 

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