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Abstrak :
Summary: The definitive "bible" for the field of biomedical engineering, this collection of volumes is a major reference for all practicing biomedical engineers and students. Now in its fourth edition, this work presents a substantial revision, with all sections updated to offer the latest research findings. New sections address drugs and devices, personalized medicine, and stem cell engineering. Also included is a historical overview as well as a special section on medical ethics. This set provides complete coverage of biomedical engineering fundamentals, medical devices and systems, computer applications in medicine, and molecular engineering

Abstrak :
Biomedical foams are a new class of materials, which are increasingly being used for tissue engineering applications. Biomedical Foams for Tissue Engineering Applications provides a comprehensive review of this new class of materials, whose structure can be engineered to meet the requirements of nutrient trafficking and cell and tissue invasion, and to tune the degradation rate and mechanical stability on the specific tissue to be repaired. Part one explores the fundamentals, properties, and modification of biomedical foams, including the optimal design and manufacture of biomedical foam pore structure for tissue engineering applications, biodegradable biomedical foam scaffolds, tailoring the pore structure of foam scaffolds for nerve regeneration, and tailoring properties of polymeric biomedical foams. Chapters in part two focus on tissue engineering applications of biomedical foams, including the use of bioactive glass foams for tissue engineering applications, bioactive glass and glass-ceramic foam scaffolds for bone tissue restoration, composite biomedical foams for engineering bone tissue, injectable biomedical foams for bone regeneration, polylactic acid (PLA) biomedical foams for tissue engineering, porous hydrogel biomedical foam scaffolds for tissue repair, and titanium biomedical foams for osseointegration.

Abstrak :
Latar Belakang: Bone tissue engineering merupakan alternatif untuk remodeling tulang pada defek kritis melalui pemanfaatan scaffold tiga dimensi berbahan polimer maupun ceramic. Bahan dari alam seperti propolis telah terbukti mampu meningkatkan pembentukan tulang baru melalui pemanfaatannya secara tunggal maupun dengan material lainnya. Namun, penambahan propolis dengan polymer-ceramic based scaffold belum pernah dilakukan sebelumnya. Tujuan: Mengevaluasi pengaruh propolis terhadap fasa kristal hidroksiapatit dan ukuran pori scaffold hidroksiapatit-gelatin-propolis melalui karakterisasi XRD dan SEM. Metode: Karakterisasi XRD dan SEM dilakukan pada scaffold dengan prosedur yang diadaptasi dari penelitian Sunarso et al. (2011). Agen crosslink pada penelitian ini menggunakan glutaraldehida. Scaffold dikarakterisasi dengan XRD untuk mengamati fasa kristal hidroksiapatit dan SEM untuk mengamati morfologi permukaan. Hasil: Pada seluruh spesimen, fasa kristal masih didominasi oleh Ca(OH)2 dan kadungan hidroksiapatit menurun seiring penambahan propolis. Semakin tinggi konsentrasi propolis, ukuran pori semakin meningkat dengan rentang rata-rata diameter pori dari seluruh spesimen 87 μm-112 μm. Kesimpulan: Hidroksiapatit pada penelitian ini tidak terbentuk sempurna dan penambahan propolis menurunkan kristalitas hidroksiapatit. Secara morfologi, spesimen yang dihasilkan memenuhi syarat scaffold. ......Background: Bone tissue engineering is an alternative for bone remodeling in critical defects through the use of three-dimensional scaffold made from polymer or ceramic. Natural material such as propolis has been shown to increase new bone formation through their use alone or with other materials. However, incorporation of propolis to polymer-ceramic based scaffold has never been done before. Objective: To evaluate the effect of propolis on hydroxyapatite crystal phase and pore size of hydroxyapatite-gelatin-propolis scaffold through XRD and SEM characterization. Methods: XRD and SEM characterization was carried out on scaffold made from chemical mixing procedure adopted from Sunarso et al. (2011). This study used glutaraldehyde as crosslink agent. The scaffold was characterized by XRD to observe the hydroxyapatite crystal phase and SEM to observe the surface morphology. Results: The crystal phase from all specimens is still dominated by Ca(OH)2 and the hydroxyapatite content is decreasing as the addition of propolis. Addition of propolis also increasing the pore size increases with the average range is 87 μm-112 μm. Conclusion: The hydroxyapatite in this study is not fully formed and the addition of propolis decreases the crystallinity of hydroxyapatite. Morphologically, all specimens fulfill the scaffold requirements.

Abstrak :
Latar Belakang: Rekonstruksi pada defek tulang kritikal masih merupakan tantangan yang besar untuk seorang ahli bedah plastik rekonstruksi. Selama ini, baku emas untuk menangani kasus defek tulang tersebut adalah menggunakan autologous bone graft, namun terdapat beberapa kekurangannya seperti morbiditas pada lokasi donor, pemanjangan waktu operasi, donor yang terbatas, dan pemajangan waktu rawat. Mencoba mengatasi kekurangan tadi, muncullah rekayasa jaringan tulang yang memberikan hasil yang menjanjikan dalam regenerasi jaringan tulang biologis yang baru. Beberapa penelitian hewan sebelum, menunjukkan bahwa implantasi secara ortotopik dan ektopik dapat memberikan hasil yang cukup baik dalam regenerasi tulang Metode: Telaah sistematis dilakukan pada Pubmed/MEDLINE, Cochrane Library, dan WHO ICTRP, termasuk semua studi dengan data primer untuk rekayasa jaringan tulang menggunakan kalsium fosfat sebagai bahan rangka, studi pada defek tulang kritikal, baik uji klinis acak terkontrol maupun tidak pada manusia dan hewan. Luaran yang dinilai adalah pembentukan tulang baru yang membandingkan implantasi secara ortotopik (intraperiosteum) dan ektopik (intramuskular). Studi ini menggunakan SYRCLE’s tools untuk menilai risiko bias studi pada hewan. Hasil: Didapatkan lima studi hewan yang memenuhi kriteria eligibilitas dari total 80 studi yang diinklusi pada telaah ini. Dicantumkan karakteristik demografis dari masing-masing studi. Studi yang memiliki luaran klinis yang sama (% area tulang dan % kontak) dibandingan antara implantasi ortotopik dan ektopik. 2 studi menunjukkan bahwa implantasi secara intramuskular menggunakan kerangka yang sudah ditambahkan BMSC memberikan hasil yang baik pada pembentukan jaringan tulang baru. Kerangka kosong tidak menunjukkan adanya pembentukan tulang. Penambahan BMP-2 sebagai factor pertumbuhan dapat meningkatkan osteogenisitas baik pada implantasi ortotopik maupun ektopik. Kesimpulan: Implantasi ortotopik dapat menginduksi pembentukan tulang baru lebih baik daripada implantasi ektopik. Menggunakan kerangka yang ditambahkan BMSC serta BMP-2 pada implantasi intramuskular memberikan hasil yang baik untuk pembentukan tulang baru. Rekayasa jaringan tulang memungkinkan untuk dilakukan dengan implantasi secara ortotopik maupun ektopik ......Background: Critical bone defect reconstruction remains a major challenge in plastic reconstructive surgery. While autologous bone graft is still considered as the gold standard for treating critical bone defects, there are disadvantages like donor site morbidity long operative time, donor limitation, and extended hospital stay. In order to resolve them, bone tissue engineering has emerged in reconstruction medical studies, for they give promising result in regenerating new biological bone tissue. Previous animal studies have shown that implantating orthotopically and ectopically gave promising result in bone regeneration. Methods: A systematic search was done on PubMed/MEDLINE, Cochrane Library, and WHO ICTRP, including all studies with primary data for bone tissue engineering using calcium phosphate as scaffold materials, studies in critical bone defects, RCT or non RCT in human studies or animal studies. Studies with outcome of new bone formation comparing orthotopic (intraperiosteum) implantation and ectopic (intramusculuar) implantation. We used SYRCLE’s tools for assessing risk of bias of animal studies. Results: Five animal studies meet the eligibility criteria from a total of 80 studies are included for this review. Characteristics demography of each study are stated. Studies with the same outcome (bone area% and contact%) are compared in orthotopic and ectopic implantation. Two studies showed that intramuscular implantation using BMSC-seeded scaffold give promising result of new bone formation. However empty scaffold did not show any bone formation. Adding BMP-2 for growth factor can improved osteogenecity both in orthotopic implantation and ectopic implantation Conclusion: Orthotopic implantation can induced new bone formation better than ectopic implantations. Using BMSC-seeded and addition of BMP-2 for intramuscular implantation give good result of new bone formation. Both orthotopic and ectopic (intramuscular) implantation are possible for bone tissue engineering

Abstrak :
This book focuses on the mechanobiological principles in tissue engineering with a particular emphasis on the multiscale aspects of the translation of mechanical forces from bioreactors down to the cellular level. The book contributes to a better understanding of the design and use of bioreactors for tissue engineering and the use of mechanical loading to optimize in vitro cell culture conditions. It covers experimental and computational approaches and the combination of both to show the benefits that computational modelling can bring to experimentalists when studying in vitro cell culture within a scaffold. With topics from multidisciplinary fields of the life sciences, medicine, and engineering, this work provides a novel approach to the use of engineering tools for the optimization of biological processes and its application to regenerative medicine. The volume is a valuable resource for researchers and graduate students studying mechanobiology and tissue engineering. For undergraduate students it also provides deep insight into tissue engineering and its use in the design of bioreactors. The book is supplemented with extensive references for all chapters to help the reader to progress through the study of each topic.

Abstrak :
Advances in genetic medicine and stem cell technology have significantly improved the potential to influence cell and tissue performance, and have expanded the field towards regenerative medicine. This book offers an overview of tissue engineering and regenerative medicine

Abstrak :
Pendahuluan: Proses degeneratif yaitu berkurangnya kemampuan sel dalam menjalankan fungsi dan kematian sel karena metabolisme tubuh yang lemah. Penyakit degeneratif terjadi pada usia manula 50 tahun. Salah satu penyakit degeneratif adalah ostearthiritis, osteoarthritis menduduki 10 besar penyebab disabilitas yang disebabkan oleh degeneratif. Kejadian osteoarthritis pada tahun 2050 meningkat 20% diseluruh dunia tidak terkecuali Indonesia mengalami kenaikan 5% pada kurun beberapa tahun. Osteoarthritis merupakan kerusakan sendi yang terjadi penuaan dikarenakan kurangnya produksi kolagen yang sulit beregenerasi, kerusakan terjadi pada jaringan osteochondral yaitu jaringan pada sendi dan pada ujung tulang yang dilapisi oleh kartilago artikular. Jaringan kartilago artikular memiliki kekurangan meregenerasi yaitu sulit memperbaiki jaringan apabila terjadi kerusakan. Metode rekayasa jaringan memberikan pilihan terbaik dengan menggunakan mesenchymal stem cells, scaffold dan senyawa kimia signaling untuk mengembalikan kerusakan tersebut. Tujuan: Fabrikasi scaffold graphene oxide /hyaluronate/fibrin yang dapat menginduksi osteogenesis pada perbaikan rekayasa jaringan osteochondral dengan mengkarakterisasi sifat scaffold dengan parameter uji fisika, kimia, dan biologi. Metode: Sintesis kimia; fabrikasi scaffold metode freeze driying; karakterisasi SEM dan FTIR; uji tekan dan porositas; uji swelling, wettability, dan laju degradasi; uji biokompabilitas (viabilitas sel kualitatif dan kuantitatif (MTS assay); uji diferensiasi sel (pewarnaan alizarin red); dan analisis statistik. Hasil: Fabrikasi scaffold dibagi menjadi tiga kelompok GO, GOHA, dan GOHAF dengan metode freeze drying diameter 1 cm dan luas permukaan 4,17 cm2. Karakterisasi uji SEM rentang ukuran pori sebesar 20 – 200 μm. Pada scaffold GO 100 – 250 μm, GOHA 80 – 200 μm, dan GOHAF 20 – 150 μm. FTIR scaffold GO terdapat gugus O-H, C=O, C=C, C-OH, dan C-H; pada scaffold GOHA terdapat gugus O-H, C=O, C=C, C-OH, C-H, dan amida II; pada scaffold GOHAF terdapat gugus O-H, C=O, C=C, C-OH, C-H, amida II dan amida I. Uji mekanik tekan pada kekerasan tekan scaffold GO sebesar 294 KPa, GOHA dan GOHAF sebesar 194 KPa. Sedangkan pada Young’s Modulus GO 0.09 MPa lebih kecil dibandingkan GOHA 0.11 MPa, dan GOHAF 0.10 MPa. Laju porositas pada GO lebih besar yaitu berturut-turut H+1 sebesar 77%, H+3 sebesar 67%, dan H+5 sebesar 61%; scaffold GOHA lebih rendah yaitu H+1 sebesar 41%, H+3 sebesar 30%, dan H+5 sebesar 18%; scaffold GOHAF lebih rendah H+1 sebesar 37%, H+3 sebesar 24%, dan H+5 sebesar 11%. Rasio swelling terbaik yaitu pada scaffold GOHAF lebih rendah 8,48%. Kapasitas wettability terbaik yaitu pada scaffold GOHAF lebih rendah 28%. Rasio laju degradasi terbaik yaitu pada scaffold GOHAF lebih rendah 0.30%. Persentase viabilitas sel kualitatif (direct) terbaik yaitu scaffold GOHAF sebesar 75% dan persentase viabilitas sel kualitatif (indirect) terbaik yaitu scaffold GOHAF sebesar 109% pada perendaman 48 jam dan 72 jam dengan nilai absorbansi 0,72 OD. Uji diferensiasi sel osteogenik yang terbaik yaitu pada scaffold GOHAF sebesar 905% terdiferensiasi menjadi sel osteogenik dengan absorbansi 0,0915 OD. Terdapat pengaruh komposisi scaffold graphene oxide/hyaluronate/fibrin (GOHAF) terhadap jumlah induksi osteogenesis atau terdiferensiasi menjadi sel osteogenik dengan hasil uji statistik signifikasi p value <0,05. ......Introduction: The degenerative processes, namely the reduced ability of cells to carry out functions and cell death due to weak metabolism. Degenerative diseases occur in seniors aged 50 years. One of the degenerative diseases is osteoarthritis, osteoarthritis occupies the top 10 causes of disability caused by degenerative. The incidence of osteoarthritis in 2050 increases by 20% worldwide, including Indonesia, which has increased by 5% in several years. Osteoarthritis is joint damage that occurs with aging due to a lack of collagen production, which is difficult to regenerate, damage occurs in osteochondral tissue, namely the tissue in the joints and at the ends of bones covered by articular cartilage. The articular cartilage tissue has the disadvantage of regenerating that it is difficult to repair the tissue if there is damage. The tissue engineering method provides the best choice by using mesenchymal stem cells, scaffold, and chemical signaling compounds to reverse the damage. Objective: Fabrication of graphene oxide/hyaluronate/fibrin scaffold for induced osteogenesis in osteochondral tissue engineering repair. Methods: Chemical synthesis; scaffold fabrication freeze driying method; SEM and FTIR characterization; compressive and porosity test; swelling, wettability, and degradation rate tests; biocompatibility test (qualitative and quantitative cell viability (MTS assay)); cell differentiation test (alizarin red stain); and statistical analysis. Result: Scaffold fabrication was divided into three groups GO, GOHA, and GOHAF by a freeze-drying method with a diameter of 1 cm and a surface area of 4.17 cm2. Characterization of SEM test pore size ranges of 20-200 m. The scaffold is GO 100 - 250 m, GOHA 80 - 200 m, and GOHAF 20 - 150 m. FTIR scaffold GO contains O-H, C = O, C = C, C-OH, and C-H groups; on the GOHA scaffold there are O-H, C = O, C = C, C-OH, C-H, and amide II groups; on the GOHAF scaffold, there are groups of O-H, C = O, C = C, C-OH, C-H, amide II and amide I. The compressive mechanical test on the compressive hardness of the GO scaffold is 294 KPa, GOHA and GOHAF are 194 KPa. Whereas in Young's Modulus GO 0.09 MPa is smaller than GOHA 0.11 MPa, and GOHAF 0.10 MPa. The porosity rate in GO was greater in H + 1 of 77%, H + 3 of 67%, and H + 5 of 61%; than GOHA scaffold was lower in H + 1 by 41%, H + 3 by 30%, and H + 5 by 18%; than GOHAF scaffold was lower in H + 1 by 37%, H + 3 by 24%, and H + 5 by 11%. The best swelling ratio is the GOHAF scaffold which is 8.48%. The best wettability capacity is the GOHAF scaffold, which is 28%. The best degradation rate ratio is the GOHAF scaffold which is 0.30%. The best qualitative (direct) cell viability percentage was 75% GOHAF scaffold and the best qualitative (indirect) cell viability percentage was 109% GOHAF scaffold immersion for 48 hours and 72 hours with an absorbance value of 0.72 OD. The best osteogenic cell differentiation test is the GOHAF scaffold, which is 905% differentiated into osteogenic cells with an absorbance of 0.0915 OD. There is an effect on the composition of the scaffold graphene oxide/hyaluronate/fibrin (GOHAF) in the amount of osteogenesis induction or differentiation into osteogenic cells with statistical test results of significance p-value <0.05.

Abstrak :
Osteoartritis merupakan penyakit kronis yang ditandai dengan kemunduran tulang rawan dan menyebabkan kekakuan, nyeri, dan gangguan pergerakan. Strategi rekayasa jaringan tulang menggunakan perancah dapat menjadi alternatif yang menjanjikan untuk regenerasi jaringan tulang yang rusak. Penelitian ini bertujuan untuk fabrikasi dan karakterisasi perancah dengan material chitosan (CS), hyaluronic acid (HA), hydroxyapatite (Hap) dengan kombinasi penambahan graphite (Gr), graphene oxide (GO), dan multiwalled carbon nanotube (MWNCT) untuk aplikasi rekayasa jaringan tulang. Dalam penelitian ini, dilakukan sintesis GO dan fungsionalisasi kimia dari material Gr dan MWNCT. Fabrikasi perancah dilakukan dengan metode freeze drying. Seluruh kelompok perancah dilakukan karakterisasi SEM dan FTIR, uji tekan dan porositas, uji swelling, wettability, dan laju degradasi. Fabrikasi perancah dibagi menjadi empat kelompok yaitu CS/HA/HAp, CS/HA/HAp/GO, CS/HA/HAp/f-Gr, dan CS/HA/HAp/f-MWNCT dengan ukuran diameter 1 cm, tinggi 1,5 cm, dan luas permukaan luas permukaan 4,71-6,28 cm2. Keseluruhan perancah memiliki ukuran pori yang bervariasi dan terdistribusi pada permukaan. Berdasarkan hasil FTIR, perancah mengandung gugus fungsi O-H, C=O, C-O-C, amida I, amida II, dan fosfat (PO43-). Pada uji kekuatan tekan, keseluruhan perancah memiliki CS/HA/HAp memiliki kekuatan tekan dan young modulus yang serupa dengan cancellous bone sebesar 5,76-6,14 MPa dan 3,95-471 MPa. Perancah memiliki laju porositas dengan rentang 13,8- 86,6%. Perancah memiliki kemampuan wettabiliy yang baik dengan rentang persentase 726-1069%. Rasio swelling perancah berada pada rentang 25,2-39,3%. Laju degradasi perancah cukup terkontrol dengan rentang 16,7-35,5%. Berdasarkan seluruh hasil karakterisitik, perancah CS/HA/HAp dengan penambahan GO merupakan kandidat terkuat sebagai perancah ideal pada penelitian ini. Perancah GO mempunyai karakteristik yang berada diantara perancah kontrol dan perancah f-MWNCT/f-Gr. ......Osteoarthritis is a chronic disease characterized by the deterioration of cartilage and causes stiffness, pain, and impaired movement. The bone tissue engineering strategy using scaffolds can be a promising alternative for the regeneration of damaged bone tissue. This study aims to fabricate and characterize scaffolds with chitosan (CS), hyaluronic acid (HA), hydroxyapatite (Hap) with a combination of addition of graphite (Gr), graphene oxide (GO), and multiwalled carbon nanotubes (MWNCT) for tissue engineering applications. In this study, GO synthesis and chemical functionalization of Gr and MWNCT materials were carried out. Scaffolding was done by freeze drying method. All groups of scaffolds were characterized by SEM and FTIR, compressive and porosity tests, swelling, wettability, and rate of degradation tests. Scaffolding was divided into four groups, namely CS/HA/HAp, CS/HA/HAp/GO, CS/HA/HAp/f-Gr, and CS/HA/HAp/f-MWNCT with a diameter of 1 cm, height 1, 5 cm, and a surface area of ​​4.71-6.28 cm2. The entire scaffold has varying pore sizes and is distributed over the surface. Based on the results of FTIR, the scaffold contains functional groups O-H, C=O, C-O-C, amide I, amide II, and phosphate (PO43-). In the compressive strength test, all scaffolds having CS/HA/HAp had similar compressive strength and young modulus with cancellous bone of 5.76-6.14 MPa and 3.95-471 MPa. Scaffolds have porosity rates in the range of 13.8-86.6%. Scaffolds have good wetability with a percentage range of 726-1069%. The swelling ratio of the scaffolds was in the range of 25.2-39.3%. The rate of degradation of the scaffold was quite controlled with a range of 16.7-35.5%. Based on all the characteristic results, the CS/HA/HAp scaffold with the addition of GO was the strongest candidate as the ideal scaffold in this study. The GO scaffold has characteristics that are between the control scaffold and the f-MWNCT/f-Gr scaffold.

Abstrak :
Sebagai salah satu organ yang berada di dalam sistem rangka tubuh manusia, tulang merupakan organ yang memiliki kemampuan untuk melakukan regenerasi mandiri. Namun, kapasitas untuk meregenerasi dapat terganggu akibat beberapa faktor seperti usia, besarnya kerusakan, dan penyakit yang diderita. Kemajuan perkembangan di bidang medis memunculkan fenomena pengaplikasian perancah rekayasa jaringan tulang untuk menjadi salah satu solusi. Dalam penelitian ini, perancah dibuat menggunakan campuran antara polyvinyl alcohol (PVA) dengan polycaprolactone (PCL). Kombinasi dilakukan dengan penambahan kandungan zirconia (ZrO2) yang memiliki kekuatan mekanik tinggi dan modifikasi permukaan lewat pelapisan gelatin. Pembuatan perancah digunakan dengan metode freeze dry dan menghasilkan lima variasi kelompok yaitu PVA/PCL, PVA/PCL berlapis gelatin, PVA/PCL/2,5%ZrO2 berlapis gelatin, PVA/PCL/5%ZrO2 berlapis gelatin, dan PVA/PCL/7,5%ZrO2 berlapis gelatin. Karakteristik fisika-kimia perancah yang terlihat antara lain perancah memiliki kekuatan tekan di rentang 4 – 19 MPa; bentuk pori perancah terbentuk di rentang 102 – 209 μm dan terbentuk porositas di rentang 67 – 83%; permukaan hidrofilik dengan tingkat swelling di rentang 224 – 452%; dan perancah memiliki laju degradasi yang cukup cepat dengan kehilangan berat di rentang 49 – 77% pada hari ketujuh. Berdasarkan karakteristik fisika-kimia, perancah mampu menyamai kekuatan tekan dan porositas pada tulang sponge. Adanya penambahan zirconia juga berhasil meningkatkan kekuatan mekanik dan memperlambat laju degradasi. Oleh karena itu, perancah PVA/PCL/ZrO2 berlapis gelatin merupakan kandidat yang baik digunakan untuk aplikasi rekayasa jaringan tulang. ......As a crucial component of the human skeletal system, bone possesses intrinsic self-regenerative capabilities. However, these regenerative capacities can be compromised by factors such as aging, the extent of injury, and the presence of certain diseases. Recent advancements in medical science have led to the development of bone tissue engineering scaffolds as a promising therapeutic solution. In this study, scaffolds were fabricated using a blend of polyvinyl alcohol (PVA) and polycaprolactone (PCL), with the addition of zirconia (ZrO2) for its high mechanical strength, and surface modification through gelatin coating. The scaffolds were produced using the freeze-drying method, resulting in five distinct groups: PVA/PCL, gelatin coated PVA/PCL, gelatin coated PVA/PCL/2,5%ZrO2, gelatin coated PVA/PCL/5%ZrO2, and gelatin coated PVA/PCL/7,5%ZrO2. The scaffolds physicochemical properties were characterized by a compressive strength ranging from 4 to 19 MPa; pore sizes between 102 and 209 μm with porosity levels from 67% to 83%; hydrophilic surfaces with swelling ratios from 224% to 452%; and a rapid degradation rate with a weight loss ranging from 49% to 77% by the seventh day. These physicochemical characteristics indicate that the scaffolds emulate the compressive strength and porosity of cancellous bone. The addition of zirconia significantly enhanced mechanical strength and decelerated the degradation rate. Consequently, gelatin coated PVA/PCL/ZrO2 scaffolds are viable candidates for applications in bone tissue engineering.