Biofunctionalization of Hydroxyapatite for craniofacial bone regeneration
- Alternative Title
- 두개안면골 재생을 위한 수산화인회석의 생체기능화
- Abstract
- Hydroxyapatite (HA) has been widely used in bone regeneration due to its excellent biocompatibility and osteoconductivity. However, poor bioactivity and lack of osteoinductivity have led to employing various tissue engineering strategies to address the issue. This thesis aims at discussing and analyzing new approaches for biofunctionalizing hydroxyapatite towards craniofacial bone regeneration. The first approach involved isolation, characterization, in vitro and in vivo testing of hydroxyapatite extracted from Parrotfish (Scardiae Collana) bone. Physico-chemical properties of the isolated HA showed it was carbonated, nano sized and rod shaped with the presence of trace elements. In vitro biological activities indicated an enhanced biocompatibility, higher alkaline phosphatase activity and mineralization ability suggestive of the osteogenic capacity of the fish-derived HA compared to synthetic HA. In vivo rabbit model testing observed significantly higher bone volume and bone mineral density in the defects filled with fishbone hydroxyapatite to synthetic hydroxyapatite.
The effect of silicon inclusion in hydroxyapatite on bone healing and osseointegration in animal models was systematically studied and evaluated. All the sixteen studies included in the analysis revealed that the addition of silicon to hydroxyapatite improved bone healing and osseointegration. Furthermore, three-dimensional printed silicon-hydroxyapatite incorporated with gelatin membrane was compared with hydroxyapatite incorporated gelatin and gelatin alone for guide bone regeneration. Physico-chemical properties reflected successful synthesis and fabrication of silicon-hydroxyapatite incorporated with gelatin membrane with comparable mechanical properties to hydroxyapatite incorporated gelatin and gelatin membranes. Improved biological interactions were observed with increased osteogenic potential in relation to silicon-hydroxyapatite membranes.
Bone regenerative capacity of the bone substitutes could be boosted by incorporating bioactive agents such as proteins, and different drugs. Simvastatin, an inhibitor of cholesterol synthesis, have demonstrated to stimulate bone formation by enhancing the expression of bone morphogenetic protein-2 (BMP-2) in osteoblasts. This study evaluated bone regeneration following simvastatin loaded nano hydroxyapatite scaffold in the bone defect created on the femoral condyle of rabbits. Quantitative and qualitative analysis exhibited significantly higher bone volume and bone mineral density in the defects filled with simvastatin loaded nano-hydroxyapatite compared to nano hydroxyapatite alone. These observations suggested that the strategies promoted bioactivity and osteoinduction resulting in an enhanced osteogenesis and bone healing.
Hydroxyapatite(HA)는 우수한 생체 적합성과 골 전도성으로 인하여 골 조직 재생에 널리 사용되고 있으나 낮은 생체 활성도 및 부족한 골 유도성으로 인한 문제가 보고되고 있다. 따라서 본 연구에서는 두개안면골 재생을 위한 HA 생체 기능화의 새로운 방법들을 논의하고 분석하고자 한다. 첫 번째 연구주제는 Parrotfish(Scardiae Collana)뼈에서 분리한 HA의 추출법과 특성분석, 세포실험 및 동물실험에 대한 내용이다. 추출된 HA의 물리 화학적 특성은 미량의 원소들로 이루어진 나노막대 형상을 한 탄산화물질임이 확인되었다. 세포실험을 통해 측정된 결과에서는 Parrotfish뼈에서 추출해 낸 HA가 합성 HA에 비해 보다 우수한 생체적합성을 보였고 골 형성능력을 나타내는 염기성 인산분해효소 및 무기질화 수치 또한 높게 측정되었다. 토끼 모델을 이용한 동물실험에서는 Parrotfish뼈에서 추출한 HA로 채워진 골 결손부가 합성 HA로 채워진 골 결손부에 비해 신생골의 형성과 골 밀도가 현저히 높게 관찰되었다.
두 번째 연구주제에서는 실리콘이 함유된 HA(Si-HA)의 뼈 치유 능력 및 골 유착능에 대한 효과가 동물실험을 통해 체계적으로 연구되고 평가되었다. 분석을 진행한 16개의 모든 실험에서 HA에 실리콘을 첨가하였을 때 뼈 치유 능력 및 골 유착능이 향상되었음이 확인되었다. 또한, 골 유도재생술에 사용 될 수 있는 Si-HA 젤라틴 합성 membrane이 3D 프린팅을 이용하여 제작되었고 HA 젤라틴 합성 membrane, 젤라틴 membrane 과 비교하여 실험되었다. 물리 화학적 특성에 대한 분석은 Si-HA 젤라틴 합성 membrane 이 HA 젤라틴 합성 membrane, 젤라틴 membrane과 유사한 기계적 특성을 나타내어 성공적으로 합성되고 제작되었다는 것을 보여주었다. 생물학적 특성분석에서는 Si-HA membrane 에 관하여 증가된 골 형성능과 함께 향상된 생물학적 상호작용들이 관찰되었다.
골 이식재의 재생능력은 단백질과 같은 생리활성제들과 혼합되면서 향상될 수 있다. 콜레스테롤 합성 억제제인 simvastatin은 조골 세포의 BMP-2 발현을 향상시켜 골 형성을 자극하는 것으로 입증되어있다. 세 번째 연구주제에서는 토끼의 대퇴골관절구에 생성한 골 결손부에서의 simvastatin이 첨가된 nano-HA 세포지지체의 골 재생을 분석하였다. 정량 및 정성분석은 simvastatin이 첨가된 nano-HA 세포지지체가 nano-HA 세포지지체보다 현저히 높은 골 형성과 골 밀도를 나타내었다. 이러한 연구들은 생체 활성 및 골 유도를 촉진하여 향상된 골 형성과 뼈 치유를 하는 결과를 얻는 새로운 방안으로 제시될 수 있을 것이라 기대된다.
- Issued Date
- 2020
- Awarded Date
- 2020. 2
- Type
- Dissertation
- Publisher
- 부경대학교
- Affiliation
- Pukyong Natiional University, Graduate School
- Department
- 대학원 의생명기계전기융합공학협동과정
- Advisor
- Seung Yun Nam
- Table Of Contents
- 1. Introduction 1
1.1. Composition of Bone 2
1.2. Structural Types of Bone 3
1.3. Cell Types: Characteristics, Functions and Properties 5
1.3.1. Osteoblasts 7
1.3.2. Osteocytes 10
1.3.3. Osteoclasts 11
1.4. Bone - Mechanical Properties 14
1.5. Bone Remodeling and Repair 15
1.6. Bone Graft 20
1.6.1. Autografts 21
1.6.2. Allografts 23
1.6.3. Xenografts 24
1.6.4. Alloplastic Grafts 25
1.7. Biomaterials 26
1.7.1. Properties of Scaffold 26
1.7.2. Bio ceramics 30
1.8. Hydroxyapatite 30
1.8.1. Synthesis of Hydroxyapatite 31
1.8.2. Physical and chemical properties of hydroxyapatite 31
1.8.3. Biological properties of Hydroxyapatite 33
1.8.4. Functionalization of hydroxyapatite 34
1.9. Aims and Objectives 37
2. Isolation and characterization of nanorod-shaped crystalline hydroxyapatite from Parrotfish bone 40
2.1. Introduction 41
2.2. Materials and Methods 43
2.2.1. Fish bone preparation 43
2.2.2. Fourier transform infrared spectroscopy (FT-IR) 44
2.2.3. X-ray diffraction (XRD) analysis 45
2.2.4. Cell proliferation assay 46
2.2.5. Morphological studies with optical microscopy and Hoechst stain 33342 46
2.2.6. Osteogenic activity 47
2.2.7. Statistical analysis 49
2.3. Results 49
2.3.1. Fourier transform infrared spectroscopy 49
2.3.2. X-ray diffraction analysis 51
2.3.3. Transmission electron microscopy analysis 52
2.3.4. Cell proliferation assay 53
2.3.5. Morphological studies with optical microscopy with Hoechst stain 33342 55
2.3.6. Osteogenic activity 55
2.4. Discussion 59
2.5. Conclusions 63
3. In Vivo Behaviors of Fishbone Hydroxyapatite for Bone Regeneration 64
3.1. Introduction 65
3.2. Materials and Methods 66
3.2.1 Material preparation 66
3.2.2. Animal Surgery 67
3.2.3. Micro Computated tomography 68
3.2.4. Histology 69
3.2.5. Statistical Analysis 69
3.3. Results 70
3.3.1. Micro Computated tomography 70
3.3.2. Histology 72
3.4. Discussion 73
3.5. Conclusions 76
4. Effect of Silicone-Doped Hydroxyapatite on Bone Regeneration and Osseointegration: A Systematic Review 78
4.1. Introduction 80
4.2. Materials and Methods 82
4.2.1. Focused question 82
4.2.2. Eligibility and exclusion criteria 83
4.2.3. Search strategy 84
4.3. Results 86
4.4. Discussion 95
4.5. Conclusion 98
5. Silicon hydroxyapatite reinforced gelatin 3D printed membrane for guided bone regeneration 99
5.1. Introduction 101
5.2. Materials and methods 103
5.2.1. Synthesis of HA and SiHA 103
5.2.2. Preparation of biomaterial 104
5.2.3 3D printing 105
5.2.4. Physico-chemical characterization 105
5.2.5. Biological characterization 106
5.2.6. Statistical analysis 109
5.3. Results and Discussion 110
5.4. Conclusion 118
6. Simvastatin Loaded Nano Hydroxyapatite in Bone Regeneration: A Study in the Rabbit Femoral Condyle 119
6.1 Introduction 121
6.2. Materials and Methods 123
6.2.1. Preparation of Simvastatin loaded nano hydroxyapatite 124
6.2.2. Surgery 124
6.2.3. Micro-computed tomography 125
6.2.4. Histology 126
6.2.5. Statistical Analysis 126
6.3. Results 127
6.3.1. Micro-computed tomography 127
6.3.2. Histology 131
6.4. Discussion 132
6.5. Conclusion 136
7.Conclusions 137
7.1. Development of bioinspired sourced HA 137
7.2. Bioactive ions addition to HA 139
7.3. Addition of growth factors 141
7.4. Publications 142
7.4.1. Journal Articles 142
7.4.2. Book Chapters 143
7.4.3. Conferences 144
7.5. Acknowledgement 145
7.6. Bibliography 148
- Degree
- Doctor
- Appears in Collections:
- 대학원 > 의생명기계전기융합공학협동과정
- Files in This Item:
-
-
Download
Biofunctionalization of Hydroxyapatite for craniofacial bone regeneration.pdf
기타 데이터 / 5.86 MB / Adobe PDF
-
Items in Repository are protected by copyright, with all rights reserved, unless otherwise indicated.