A study on effects of hydroxyapatite nanoparticles and calcium carbonate microcapsules on properties and biocompatibility of a thermosensitive chitosan/collagen hydrogel

การศึกษาผลของนาโนไฮดรอกซีแอปาไทต์และแคลเซียมคาร์บอเนตไมโครแคปซูลต่อคุณสมบัติและความเข้ากันได้ทางชีวภาพของไคโตซาน/คอลลาเจน ไฮโดรเจลที่มีความไวต่ออุณหภูมิ

Name: Natthaporn Jaiman

keyword: Biocompatibility

; Bone morphogenetic protein-2

; Calcium carbonate microcapsules

; Ectopic bone formation

; Hydroxyapatite nanoparticles

; Thermosensitive chitosan/collagen hydrogel

Abstract: To develop thermosensitive chitosan/collagen hydrogels that are osteoconductive and injectable for alveolar bone regeneration. The current study aims to improve the properties of hydrogel by adjusting ratio of calcium carbonate (CaCO3) microcapsules and nanohydroxyapatite particles (nHA) in the beta glycerophosphate chitosan/collagen hydrogel. Material & method: There were 2 parts of study. Part I: Hydrogel characterizations, there are two subsets for the nHA and CaCO3 microcapsule sets (Groups I-IV) as followed, nHA set: nHA 2% and nHA 5% (w/v), and CaCO3 set: Group I: CaCO3 0%, Group II: CaCO3 1%, Group III: CaCO3 2%, and Group IV: CaCO3 5% (w/v). Including microstructures, mechanical properties, physical properties, and cytotoxicity test. Part II: Subcutaneous implants in mice for histology analysis to evaluate biocompatibility and new bone formation, there were four groups of study (Groups A-D), Group A: Calcium carbonate microcapsules, Group B: thermosensitive CaCO3-nHA-chitosan/collagen hydrogel, Group C: thermosensitive CaCO3-nHAchitosan/collagen hydrogel with BMP-2, and Group D: sham operation. The CaCO3 were prepared by precipitating sodium carbonate that with calcium chloride in the carboxymethyl cellulose solution, then morphology of the microcapsules was examined using scanning electron microscope (SEM). After that the thermosensitive 0%, 1%, 2%, and 5% (w/v) CaCO3-5 2% and 5% (w/v) nHA-chitosan/collagen hydrogels were prepared and the 10% (w/v) betaglycerophosphate was used as an accelerator for the sol-gel transition under the temperature change at 37ºC. After that, the effects of the 0% - 5% CaCO3 and 2% and 5% nHA on the microstructures, mechanical, and physical strengths of the hydrogels were investigated. Cell viability assay was performed on the human osteoblast cell line (hFOB1.19, ATTC, USA) that were cultured with the culture mediums pre-incubated with the hydrogels. The 2% CaCO3- 2% nHA-chitosan/collagen hydrogel with and without BMP-2 were implanted in subcutaneous pockets of mouse for 4 weeks. Then histological examination of the hematoxylin eosin and Masson trichrome staining was performed to determine levels of foreign body reaction and ectopic bone formation. The significant differences were set at p<0.05 (n=3-5, Mean ± SD). Results: It was found that CaCO3 microcapsules were 2-5 μm round shaped particles. The 5% nHA was strong particulate gel that supports distribution of the microcapsule in a threedimensional matrix of the hydrogels. The CaCO3 microcapsules increased mechanical and physical strength, while decreased porosity and swelling and degradation rates of the hydrogels. The CaCO3-nHA hydrogels were non-cytotoxic demonstrated by high levels of cell viability of cells incubated in the culture medium pre-incubated with the hydrogels. The microcapsules and hydrogels were biocompatible by stimulating mild foreign body tissue reaction and the inflammatory responses in the sample groups were comparable to the sham group. Moreover, in a group of the hydrogel with BMP-2, the patches of the newly formed bone or mineralized tissue were identified. The patches exhibited light pink stain of haematoxylin and eosin staining and blue staining of Masson trichrome staining with multiple lacunae with nuclei on the matrixes of the hydrogel with BMP-2 group. Conclusion: Properties of the hydrogel could be modified by adjusting ratios of the CaCO3 microcapsules and nHA. The 2% CaCO3 with 2% nHA-chitosan/collagen hydrogels exhibited mechanical and physical strength superior and suitable than other groups for promoting bone regeneration and could function as a delivery vehicle for BMP-2. The hydrogels were noncytotoxic and bioinert

Thammasat University. Thammasat University Library

Address: BANGKOK

Email: preserv@tu.ac.th

Name: Premjit Arpornmaeklong

Role: advisor

Created: 2022

Modified: 2024-07-16

Issued: 2024-07-16

วิทยานิพนธ์/Thesis

application/pdf

DOI: 10.14457/TU.the.2022.1472

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Dentistry

Grantor: Thammasat University

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