Biomaterials For Bone Tissue Engineering Research Articles

Scaffold Fabrication Techniques of Biomaterials for Bone Tissue Engineering: A Critical Review.

Bone tissue engineering (BTE) is a promising alternative to repair bone defects using biomaterial scaffolds, cells, and growth factors to attain satisfactory outcomes. This review targets the fabrication of bone scaffolds, such as the conventional and electrohydrodynamic techniques, for the treatment of bone defects as an alternative to autograft, allograft, and xenograft sources. Additionally, the modern approaches to fabricating bone constructs by additive manufacturing, injection molding, microsphere-based sintering, and 4D printing techniques, providing a favorable environment for bone regeneration, function, and viability, are thoroughly discussed. The polymers used, fabrication methods, advantages, and limitations in bone tissue engineering application are also emphasized. This review also provides a future outlook regarding the potential of BTE as well as its possibilities in clinical trials.

Synthesized bioactive lignin nanoparticles/polycaprolactone nanofibers: A novel nanobiocomposite for bone tissue engineering

The use of artificial biomaterial with enhanced bioactivity for osteostimulation is a major research concern at present days. In this research, antibacterial and osteostimulative core-shell lignin nanoparticles (LgNP) were synthesized from alkali lignin using tetrahydrofuran (THF) as solvent via a simultaneous pH and solvent shifting technology. Later, LgNP-loaded polycaprolactone (PCL) composite nanofibers were fabricated via the electrospinning technique. The addition of LgNP significantly increased the diameter of the nanofibers, ranging from 400 to 2200 nm. The addition of LgNP reduced the mechanical performance, crystallinity, and porosity of the nanofibers while improving surface wetting and swelling properties of the inherently hydrophobic PCL polymer. The prepared nanofibers showed excellent bactericidal efficacy against major bone infectious Gram-positive Staphylococcus aureus bacterial strains. The incorporation of LgNP imparted superior antioxidant activity and boosted the biodegradation process of the nanofibers. The deposition of biomineral apatite with platelet-like clustered protrusions having a Ca/P ratio of 1.67 was observed while incubating the scaffold in simulated body fluid. Based on the results of the LDH and WST-1 assay, it was demonstrated that the composite nanofibers are non-toxic to pre-osteoblastic cell line (MC3T3-E1) when they are placed in direct contact with the LgNP/PCL scaffold nanofibers. The MC3T3-E1 cells exhibited excellent proliferation and attachment on the prepared composite scaffold via filopodial and lamellipodial expansion with cell-secreted Ca deposition. According to the alkaline phosphatase activity test, LgNP/PCL nanofiber scaffolds significantly improved osteogenic differentiation of MC3T3-E1 cells compared to neat PCL nanofibers. Overall, our findings suggest that LgNP/PCL nanofiber scaffold could be a promising functional biomaterial for bone tissue engineering.

Zeolites: A series of promising biomaterials in bone tissue engineering.

As an important worldwide medical issue, bone defect exhibits a variety of physical and psychological consequences on sufferers. Some features of clinical treatments including bone grafting and limb shortening are not satisfactory. Recently, bone tissue engineering has been considered as the most effective approach to dealing with the issue of bone deformities. Meanwhile, a variety of biomaterials have been rationally designed and created for the bone regeneration and tissue repairing. Among all these admirable biomaterials for bone remodeling, zeolite-based materials can serve as efficient scaffold candidates with excellent osteo-inductivity. In addition, the porous nature and high biocompatibility of zeolites endow them with the ability as ideal substrates for cell adhesion and proliferation. More importantly, zeolites are investigated as potential coating materials for implants because they have been proven to increase osteo-conductivity and aid in local elastic modeling. Last but not least, zeolites can also be used to treat bone disorders and act as dietary supplements during the practical applications. Accordingly, numerous benefits of zeolite prompt us to summarize their recent biomedical progress including but not limited to the distinguishing characteristics, broad classifications, as well as promising usages in bone tissue engineering.

Cytocompatibility of MG-63 osteosarcoma cells on chitosan/hydroxyapatite/lignin hybrid composite scaffold in vitro

This study aims at fabricating promising cytocompatible hybrid biocomposite scaffolds from chitosan (CS), hydroxyapatite (HAP) and lignin (L) for bone tissue engineering by using freeze-drying technique. Different ratios of HAP to L (50:0, 37.5:12.5, 25:25 and 12.5:37.5) were taken to determine the optimum ratio for obtaining a composite with superior properties. The mechanical and biological properties of the resulting composites were investigated. The mechanical results showed that the prepared composite with a ratio of 25:25 of HAP/L exhibited a remarkable enhancement in the mechanical properties compared to the others. Additionally, it was found from the in vitro results that the addition of L enhanced the water uptake value of the resulting scaffolds indicating their increased hydrophilicity. As a result, a significant increase in the attachment and proliferation of MG-63 cell line (osteoblast like cells) was observed in composite scaffolds with L over the scaffold without L (CS/HAP). From these results, it could be suggested that the prepared composite scaffold with 25:25 of HAP/L is very promising biomaterials in bone tissue-engineering as it exhibited a better mechanical and biological properties than the other prepared composites.

Novel Approaches and Biomaterials for Bone Tissue Engineering: A Focus on Silk Fibroin.

Bone tissue engineering (BTE) represents a multidisciplinary research field involving many aspects of biology, engineering, material science, clinical medicine and genetics to create biological substitutes to promote bone regeneration. The definition of the most appropriate biomaterials and structures for BTE is still a challenge for researchers, aiming at simultaneously combining different features such as tissue generation properties, biocompatibility, porosity and mechanical strength. In this scenario, among the biomaterials for BTE, silk fibroin represents a valuable option for the development of functional devices because of its unique biological properties and the multiple chances of processing. This review article aims at providing the reader with a general overview of the most recent progresses in bone tissue engineering in terms of approaches and materials with a special focus on silk fibroin and the related mechanisms involved in bone regeneration, and presenting interesting results obtained by different research groups, which assessed the great potential of this protein for bone tissue engineering.

Effects of Substitution Ratios of Zinc-Substituted Hydroxyapatite on Adsorption and Desorption Behaviors of Bone Morphogenetic Protein-2.

Understanding interactions between bone morphogenetic proteins (BMPs) and biomaterials is of great significance in preserving the structure and bioactivity of BMPs when utilized in clinical applications. Currently, bone morphogenetic protein-2 (BMP-2) is one of the most important growth factors in bone tissue engineering; however, atomistic interactions between BMP-2 and zinc-substituted hydroxyapatite (Zn-HAP, commonly used in artificial bone implants) have not been well clarified until now. Thus, in this work, the interaction energies, binding/debinding states, and molecular structures of BMP-2 upon a series of Zn-HAP surfaces (Zn-HAPs, 1 at%, 2.5 at%, 5 at%, and 10 at% substitution) were investigated by hybrid molecular dynamics (MD) and steered molecular dynamics (SMD) simulations. Meanwhile, cellular studies including alkaline phosphatase (ALP) activity and reverse transcription-polymerase chain reaction (RT-PCR) assay were performed to verify the theoretical modeling findings. It was found that, compared to pure HAP, Zn-HAPs exhibited a higher binding affinity of BMP-2 at the adsorption process; meanwhile, the detachment of BMP-2 upon Zn-HAPs was more difficult at the desorption process. In addition, molecular structures of BMP-2 could be well stabilized upon Zn-HAPs, especially for Zn10-HAP (with a 10 at% substitution), which showed both the higher stability of cystine-knots and less change in the secondary structures of BMP-2 than those upon HAP. Cellular studies confirmed that higher ALP activity and osteogenic marker gene expression were achieved upon BMP-2/Zn-HAPs than those upon BMP-2/HAP. These findings verified that Zn-HAPs favor the adsorption of BMP-2 and leverage the bioactivity of BMP-2. Together, this work clarified the interaction mechanisms between BMP-2 and Zn-HAPs at the atom level, which could provide new molecular-level insights into the design of BMP-2-loaded biomaterials for bone tissue engineering.

A dual-responsive polydopamine-modified hydroxybutyl chitosan hydrogel for sequential regulation of bone regeneration

Delayed inflammatory reaction and poor osteogenesis are the two main causes of failure for bone-defect healing. Accordingly, in the present study, a dual-responsive hydrogel composite was successfully fabricated in which near-infrared (NIR)-light-responsive polydopamine-coated magnesium‐calcium carbonate microspheres are incorporated into a thermo-responsive hydroxybutyl chitosan hydrogel to provide sequential delivery of the anti-inflammatory drug aspirin and osteogenic bone morphogenetic protein 2 (BMP-2). By initially releasing aspirin rapidly, the hydrogel composite efficiently ameliorates early-stage inflammatory reaction and promotes transition to the regenerative phase. Then, the hydrogel composite allows NIR-light-responsive release of BMP-2, which maximizes its osteoinductive effects. Using an SD rat calvaria-defect model, the sequential and controllable release achieved by the hydrogel is demonstrated to promote new-bone formation. Thus, the current study provides an efficient alternative strategy for developing multifunctional therapeutic biomaterials for bone tissue engineering.

THE NATURAL AND COMMERCIAL SOURCES OF HYDROXYAPATITE/COLLAGEN COMPOSITES FOR BIOMEDICAL APPLICATIONS: A REVIEW STUDY

Bone is considered the core unit that forms the human body’s skeleton, consisting primarily of hydroxyapatite (HA) and collagen (Col). The composites of hydroxyapatite/collagen had been prepared through different fabricated techniques and were used in many bone defects as biomaterials for bone tissue engineering. The incorporation of HA and collagen is possible due to the biocompatibility of collagen and the high mechanical properties of the HA. HA/Col composites have been used in many medical and biological fields. Current study have been discussed the synthesis and characterization techniques of HA/Col composites; the study have been included to study the cytotoxicity and cell attachment of the composites, along with their applications, as well as barriers that still remain to their successful development for clinical application.

Strontium Functionalized in Biomaterials for Bone Tissue Engineering: A Prominent Role in Osteoimmunomodulation.

With the development of bone tissue engineering bio-scaffold materials by adding metallic ions to improve bone healing have been extensively explored in the past decades. Strontium a non-radioactive element, as an essential osteophilic trace element for the human body, has received widespread attention in the medical field due to its superior biological properties of inhibiting bone resorption and promoting osteogenesis. As the concept of osteoimmunology developed, the design of orthopedic biomaterials has gradually shifted from “immune-friendly” to “immunomodulatory” with the aim of promoting bone healing by modulating the immune microenvironment through implanted biomaterials. The process of bone healing can be regarded as an immune-induced procedure in which immune cells can target the effector cells such as macrophages, neutrophils, osteocytes, and osteoprogenitor cells through paracrine mechanisms, affecting pathological alveolar bone resorption and physiological bone regeneration. As a kind of crucial immune cell, macrophages play a critical role in the early period of wound repair and host defense after biomaterial implantation. Despite Sr-doped biomaterials being increasingly investigated, how extracellular Sr2+ guides the organism toward favorable osteogenesis by modulating macrophages in the bone tissue microenvironment has rarely been studied. This review focuses on recent knowledge that the trace element Sr regulates bone regeneration mechanisms through the regulation of macrophage polarization, which is significant for the future development of Sr-doped bone repair materials. We will also summarize the primary mechanism of Sr2+ in bone, including calcium-sensing receptor (CaSR) and osteogenesis-related signaling pathways.

Application Progress of Modified Chitosan and Its Composite Biomaterials for Bone Tissue Engineering.

In recent years, bone tissue engineering (BTE), as a multidisciplinary field, has shown considerable promise in replacing traditional treatment modalities (i.e., autografts, allografts, and xenografts). Since bone is such a complex and dynamic structure, the construction of bone tissue composite materials has become an attractive strategy to guide bone growth and regeneration. Chitosan and its derivatives have been promising vehicles for BTE owing to their unique physical and chemical properties. With intrinsic physicochemical characteristics and closeness to the extracellular matrix of bones, chitosan-based composite scaffolds have been proved to be a promising candidate for providing successful bone regeneration and defect repair capacity. Advances in chitosan-based scaffolds for BTE have produced efficient and efficacious bio-properties via material structural design and different modifications. Efforts have been put into the modification of chitosan to overcome its limitations, including insolubility in water, faster depolymerization in the body, and blood incompatibility. Herein, we discuss the various modification methods of chitosan that expand its fields of application, which would pave the way for future applied research in biomedical innovation and regenerative medicine.

A Critical Review of Additive Manufacturing Techniques and Associated Biomaterials Used in Bone Tissue Engineering.

With the ability to fabricate complex structures while meeting individual needs, additive manufacturing (AM) offers unprecedented opportunities for bone tissue engineering in the biomedical field. However, traditional metal implants have many adverse effects due to their poor integration with host tissues, and therefore new material implants with porous structures are gradually being developed that are suitable for clinical medical applications. From the perspectives of additive manufacturing technology and materials, this article discusses a suitable manufacturing process for ideal materials for biological bone tissue engineering. It begins with a review of the methods and applicable materials in existing additive manufacturing technologies and their applications in biomedicine, introducing the advantages and disadvantages of various AM technologies. The properties of materials including metals and polymers, commonly used AM technologies, recent developments, and their applications in bone tissue engineering are discussed in detail and summarized. In addition, the main challenges for different metallic and polymer materials, such as biodegradability, anisotropy, growth factors to promote the osteogenic capacity, and enhancement of mechanical properties are also introduced. Finally, the development prospects for AM technologies and biomaterials in bone tissue engineering are considered.

Bioinspired laminated bioceramics with high toughness for bone tissue engineering.

For the research of biomaterials in bone tissue engineering, it is still a challenge to fabricate bioceramics that overcome brittleness while maintaining the great biological performance. Here, inspired by the toughness of natural materials with hierarchical laminated structure, we presented a directional assembly-sintering approach to fabricate laminated MXene/calcium silicate-based (L-M/CS) bioceramics. Benefiting from the orderly laminated structure, the L-M/CS bioceramics exhibited significantly enhanced toughness (2.23 MPa·m1/2) and high flexural strength (145 MPa), which were close to the mechanical properties of cortical bone. Furthermore, the L-M/CS bioceramics possessed more suitable degradability than traditional CaSiO3 bioceramics due to the newly formed CaTiSiO5 after sintering. Moreover, the L-M/CS bioceramics showed good biocompatibility and could stimulate the expression of osteogenesis-related genes. The mechanism of promoting osteogenic differentiation had been shown to be related to the Wnt signaling pathway. This work not only fabricated calcium silicate-based bioceramics with excellent mechanical and biological properties for bone tissue engineering but also provided a strategy for the combination of bionics and bioceramics.

Current and future perspectives on biomaterials for segmental mandibular defect repair

Segmental mandibular defects are a common problem in head and neck surgery. Osseous free flap transfer has revolutionized mandibular reconstruction; however, donor site morbidity remains a significant problem. Bone tissue engineering (BTE) employs strategies for reconstructing critical-sized bone defects, improving the patients’ quality of life. Scaffolds employed for BTE must be mechanically stable, osseointegrate with neighboring bone, and promote osteogenesis. The need for modified, hybrid constructs with a diversity of favorable characteristics is essential. This review focuses on the recent progresses in modulating the physiochemical properties and applications of biomaterials in BTE in the context of mandibular segmental defect repair.

Strontium Functionalization of Biomaterials for Bone Tissue Engineering Purposes: A Biological Point of View.

Strontium (Sr) is a trace element taken with nutrition and found in bone in close connection to native hydroxyapatite. Sr is involved in a dual mechanism of coupling the stimulation of bone formation with the inhibition of bone resorption, as reported in the literature. Interest in studying Sr has increased in the last decades due to the development of strontium ranelate (SrRan), an orally active agent acting as an anti-osteoporosis drug. However, the use of SrRan was subjected to some limitations starting from 2014 due to its negative side effects on the cardiac safety of patients. In this scenario, an interesting perspective for the administration of Sr is the introduction of Sr ions in biomaterials for bone tissue engineering (BTE) applications. This strategy has attracted attention thanks to its positive effects on bone formation, alongside the reduction of osteoclast activity, proven by in vitro and in vivo studies. The purpose of this review is to go through the classes of biomaterials most commonly used in BTE and functionalized with Sr, i.e., calcium phosphate ceramics, bioactive glasses, metal-based materials, and polymers. The works discussed in this review were selected as representative for each type of the above-mentioned categories, and the biological evaluation in vitro and/or in vivo was the main criterion for selection. The encouraging results collected from the in vitro and in vivo biological evaluations are outlined to highlight the potential applications of materials’ functionalization with Sr as an osteopromoting dopant in BTE.

Homology of selenium (Se) and tellurium (Te) endow the functional similarity of Se-doped and Te-doped mesoporous bioactive glass nanoparticles in bone tissue engineering

The focus of bone tissue engineering is to realize the regeneration of new functional bone through the synergistic combination of biomaterials, therapeutic agents and cells. Doping of mesoporous bioactive glass (MBG) nanoparticles with therapeutic ions to give them special properties is gaining increasing interest in the design of biomaterials for bone tissue engineering. In this study, we synthesized Se-doped and Te-doped MBG nanoparticles using the sol-gel method, and demonstrated for the first time that the homology of Se and Te endows the functional similarity of bone tissue engineering, and also obtains the desired properties by guiding cell behavior and changing the physicochemical properties of the biomaterial. Results found that MBG nanoparticles doped with Se and Te respectively can be gained similar structure, and thus endowed their similar properties as expected, such as drug sustained release, anticancer and antibacterial properties in a dose-dependent manner. This study provides a feasible strategy for the development of homologous group ions doped nanobiomaterials and their evaluation and basic research in bone tissue engineering. Highlights • Homology of Se and Te endow the functional similarity of Se-doped and Te-doped MBGs. • Se doped MBGs exhibited excellent antibacterial activity on S. aureus (92.54%). • Te doped MBGs exhibited controllable DOX release in vitro. • Se-doped and Te-doped MBGs all exhibited high anti-bone cancer activity in vitro.

Independent effects of structural optimization and resveratrol functionalization on extracellular matrix scaffolds for bone regeneration

Due to their natural biological activity and low immunogenicity, decellularized extracellular matrix (ECM) materials have aroused interest as potential scaffold materials in tissue engineering. Decellularized small intestinal submucosa (SIS) is one ECM biomaterial that can be easily sourced. In the present study, we tested whether the osteogenesis of SIS scaffolds was enhanced via structural optimization and resveratrol (RSV) functionalization and explored the independent effects of these modifications. We obtained SIS scaffolds with different pore structures by controlling the preparation concentration. The group with superior osteogenic properties was further RSV-functionalized via covalent immobilization. We conducted a series of in vitro and in vivo studies to explore the effects of these two optimization strategies on the osteogenic properties of SIS scaffolds. The results showed that pore structure and RSV functionalization significantly affected the osteogenic properties of SIS scaffolds. With a fabrication concentration of 1%, the SIS scaffolds had superior osteogenic properties. Through covalent coupling, RSV was successfully grafted onto SIS scaffolds, where it was slowly released. The most significant improvements in osteogenic properties were obtained with a coupling concentration of 1%. Furthermore, in in vivo experiments, vascular and new bone tissue formation was enhanced with RSV/SIS scaffolds compared with SIS scaffolds and the blank control group at 4 weeks after implantation. These findings indicate that the RSV/SIS scaffolds obtained via dual optimization strategies show promise as biomaterials in bone tissue engineering.

Protocol of Co-Culture of Human Osteoblasts and Osteoclasts to Test Biomaterials for Bone Tissue Engineering.

New biomaterials and scaffolds for bone tissue engineering (BTE) applications require to be tested in a bone microenvironment reliable model. On this assumption, the in vitro laboratory protocols with bone cells represent worthy experimental systems improving our knowledge about bone homeostasis, reducing the costs of experimentation. To this day, several models of the bone microenvironment are reported in the literature, but few delineate a protocol for testing new biomaterials using bone cells. Herein we propose a clear protocol to set up an indirect co-culture system of human-derived osteoblasts and osteoclast precursors, providing well-defined criteria such as the cell seeding density, cell:cell ratio, the culture medium, and the proofs of differentiation. The material to be tested may be easily introduced in the system and the cell response analyzed. The physical separation of osteoblasts and osteoclasts allows distinguishing the effects of the material onto the two cell types and to evaluate the correlation between material and cell behavior, cell morphology, and adhesion. The whole protocol requires about 4 to 6 weeks with an intermediate level of expertise. The system is an in vitro model of the bone remodeling system useful in testing innovative materials for bone regeneration, and potentially exploitable in different application fields. The use of human primary cells represents a close replica of the bone cell cooperation in vivo and may be employed as a feasible system to test materials and scaffolds for bone substitution and regeneration.

Injectable Organic-Inorganic Biocomposites for Bone Tissue Regeneration - A Mini Review

Bone regeneration is complex physiological process, which include the most common form of regeneration - bone fracture healing and new bone formation. Moreover, large bone defects, infections and bone diseases such as osteoporosis and arthritis can impair bone regeneration. Despite intensive research and development of biomaterials for bone tissue engineering, especially for osteoporotic bone healing, the properties of the fabricated biomaterials are still far from those of unique composite structure of natural bone and desired therapeutic effect not achieved. This mini-review will highlight the various cutting-edge injectable inorganic-organic biocomposites as minimally invasive and regenerative therapeutics for bone tissue regeneration. The review will summarize the main strategic tools for the development of injectable biocomposites: natural or synthetic biopolymer-based hydrogels, bioactive inorganic fillers and biologically active components, as well as the fabrication techniques and synthesis methods.

Hydroxypropylmethyl cellulose (HPMC) crosslinked keratin/hydroxyapatite (HA) scaffold fabrication, characterization and in vitro biocompatibility assessment as a bone graft for alveolar bone regeneration

Wool derived keratin has garnered significant advancements in the field of biomaterials for hard tissue regeneration. The main limitation of keratin-based biomaterials for bone tissue engineering is their fragile nature. This paper proposes the development of a novel hydroxypropyl methylcellulose (HPMC) crosslinked keratin scaffold, containing hydroxyapatite as a major inorganic component by freeze drying technique for alveolar bone regeneration. The prepared keratin/hydroxyapatite/HPMC (K/HA/HPMC) scaffold was characterized to study its chemical, physical, and mechanical properties by Scanning electron microscope (SEM), Fourier transform infrared spectroscopy (FTIR), Energy dispersive X-ray spectroscopy (EDX), X-Ray diffractometric (XRD) analysis. The SEM images of the scaffolds showed highly porous interconnected architecture with average pore size of 108.36 ± 22.56 while microcomputed tomographic analysis measured total porosity as 79.65 %±. Energy dispersive X-ray spectroscopic (EDX) analysis confirmed that inorganic component of scaffold was mainly composed of calcium and phosphorous ions having Ca/P molar ration of 1.6. The maximum compressive strength was found to be in the range of 0.841 ± 0.37 MPa. Furthermore, the K/HA/HPMC scaffold was structurally stable and weight loss of about 26% was observed when soaked in phosphate buffered solution (PBS) for 28 days. In vitro biocompatibility testing showed that K/HA/HPMC scaffold was cytocompatible and supported the attachment, proliferation of osteoblast (Saos-2) cells. Thus, the development of a non-toxic chemical cross-linking system with HPMC was investigated to fabricate K/HA/HPMC scaffold and our results showed great potential of these scaffolds to regenerate alveolar bone due to their structural similarity and excellent in vitro biocompatibility.

Processing of biomaterials for bone tissue engineering: State of the art

Bones are the functional tissue, to provide structural support, protect vital organs, store minerals, helps in multi-directional motion and bear load of our body. Fracture of bones, in which continuity of bone tissue breaks, which occurred due to trauma (high intensity impact) and over stressing are treated with help of bone implants. Bone implants are fabricated through biomaterials which are categorized as as: (a) first generation, (b) second generation and (c) third generation biomaterials. First generation implants are bioinert, second generation implants are biodegradable, bioinert and bioactive. Third generation implants focuses on stimulating specific cellular response at molecular level along with the properties of second generation. Bone tissue engineering (BTE) is responsive for the evolution of third generation implants, where the main focus is to create the environment which helps to stimulate the cell response in direction of bone regeneration. This aim is achieved with engineered tissue implant which includes: (a) scaffold for architectural support & cell homing, (b) osteogenic cells and (c) signaling growth factors. Tissues in human body are the 3D repeating unit cells also called as extra cellular matrix (ECM). Similarly scaffold is the artificial 3D porous structure of biomaterials (ceramics, metals and polymers) which mimic those 3D repeating unit cells or ECM. Scaffold is a backbone or fundamental unit of BTE. Success of scaffold decides the success of engineered bone tissue implant. This review summarizes the factor influencing BTE, factor involved in selection of material for scaffold fabrication and trending fabrication methods for BTE scaffolds.