Mechanical properties, and in vitro biocompatibility assessment of biomimetic dual layered keratin/ hydroxyapatite scaffolds.

A novel biomimetic dual layered keratin/hydroxyapatite (keratin/HA) scaffold was designed using iterative freeze-drying technique. The prepared scaffolds were studied using several analytical techniques to better understand the biological, structural, and mechanical properties. The developed multilayered, interconnected, porous keratin scaffold with different hydroxyapatite (HA) content in the outer and inner layer, mimics the inherent gradient structure of alveolar bone. SEM studies showed an interconnected porous architecture of the prepared scaffolds with seamless integration between the upper and lower layers. The incorporation of HA improved the mechanical properties keratin/HA scaffolds. The keratin/HA scaffolds exhibited superior mechanical properties in terms of Young's modulus and compressive strength in comparison to pure keratin scaffolds. The biocompatibility studies suggested that both keratin and keratin/HA scaffolds were cyto-compatible, in terms of cell proliferation. Furthermore, it showed that both the tested materials can served as an ideal substrate for the differentiation of Saos-2 cells, leading to mineralization of the extracellular matrix. In summary, ionic liquid based green technique was employed for keratin extraction to fabricate keratin/HA scaffolds and our detailed in vitro investigations suggest the great potential for these composite scaffolds for bone tissue engineering in future.

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.

Keratin - Based materials for biomedical applications.

Keratin constitutes the major component of the feather, hair, hooves, horns, and wool represents a group of biological material having high cysteine content (7-13%) as compared to other structural proteins. Keratin -based biomaterials have been investigated extensively over the past few decades due to their intrinsic biological properties and excellent biocompatibility. Unlike other natural polymers such as starch, collagen, chitosan, the complex three-dimensional structure of keratin requires the use of harsh chemical conditions for their dissolution and extraction. The most commonly used methods for keratin extraction are oxidation, reduction, steam explosion, microbial method, microwave irradiation and use of ionic liquids. Keratin -based materials have been used extensively for various biomedical applications such as drug delivery, wound healing, tissue engineering. This review covers the structure, properties, history of keratin research, methods of extraction and some recent advancements related to the use of keratin derived biomaterials in the form of a 3-D scaffold, films, fibers, and hydrogels.