Vladimir Kasyanov

Periostin regulates collagen fibrillogenesis and the biomechanical properties of connective tissues

Periostin is predominantly expressed in collagen‐rich fibrous connective tissues that are subjected to constant mechanical stresses including: heart valves, tendons, perichondrium, cornea, and the periodontal ligament (PDL). Based on these data we hypothesize that periostin can regulate collagen I fibrillogenesis and thereby affect the biomechanical properties of connective tissues. Immunoprecipitation and immunogold transmission electron microscopy experiments demonstrate that periostin is capable of directly interacting with collagen I. To analyze the potential role of periostin in collagen I fibrillogenesis, gene targeted mice were generated. Transmission electron microscopy and morphometric analyses demonstrated reduced collagen fibril diameters in skin dermis of periostin knockout mice, an indication of aberrant collagen I fibrillogenesis. In addition, differential scanning calorimetry (DSC) demonstrated a lower collagen denaturing temperature in periostin knockout mice, reflecting a reduced level of collagen cross‐linking. Functional biomechanical properties of periostin null skin specimens and atrioventricular (AV) valve explant experiments provided direct evidence of the role that periostin plays in regulating the viscoelastic properties of connective tissues. Collectively, these data demonstrate for the first time that periostin can regulate collagen I fibrillogenesis and thereby serves as an important mediator of the biomechanical properties of fibrous connective tissues. J. Cell. Biochem. 101: 695–711, 2007.

Organ printing: from bioprinter to organ biofabrication line

Organ printing, or the layer by layer additive robotic biofabrication of functional three-dimensional tissue and organ constructs using self-assembling tissue spheroid building blocks, is a rapidly emerging technology that promises to transform tissue engineering into a commercially successful biomedical industry. It is increasingly obvious that similar well-established industries implement automated robotic systems on the path to commercial translation and economic success. The use of robotic bioprinters alone however is not sufficient for the development of large industrial scale organ biofabrication. The design and development of a fully integrated organ biofabrication line is imperative for the commercial translation of organ printing technology. This paper presents recent progress and challenges in the development of the essential components of an organ biofabrication line.

Organ printing: promises and challenges

Organ printing or biomedical application of rapid prototyping, also defined as additive layer-by-layer biomanufacturing, is an emerging transforming technology that has potential for surpassing traditional solid scaffold-based tissue engineering. Organ printing has certain advantages: it is an automated approach that offers a pathway for scalable reproducible mass production of tissue engineered products; it allows a precised simultaneous 3D positioning of several cell types; it enables creation tissue with a high level of cell density; it can solve the problem of vascularization in thick tissue constructs; finally, organ printing can be done in situ. The ultimate goal of organ-printing technology is to fabricate 3D vascularized functional living human organs suitable for clinical implantation. The main practical outcomes of organ-printing technology are industrial scalable robotic biofabrication of complex human tissues and organs, automated tissue-based in vitro assays for clinical diagnostics, drug discovery and drug toxicity, and complex in vitro models of human diseases. This article describes conceptual framework and recent developments in organ-printing technology, outlines main technological barriers and challenges, and presents potential future practical applications.

Towards organ printing: engineering an intra-organ branched vascular tree

Importance of the field: Effective vascularization of thick three-dimensional engineered tissue constructs is a problem in tissue engineering. As in native organs, a tissue-engineered intra-organ vascular tree must be comprised of a network of hierarchically branched vascular segments. Despite this requirement, current tissue-engineering efforts are still focused predominantly on engineering either large-diameter macrovessels or microvascular networks. Areas covered in this review: We present the emerging concept of organ printing or robotic additive biofabrication of an intra-organ branched vascular tree, based on the ability of vascular tissue spheroids to undergo self-assembly. What the reader will gain: The feasibility and challenges of this robotic biofabrication approach to intra-organ vascularization for tissue engineering based on organ-printing technology using self-assembling vascular tissue spheroids including clinically relevantly vascular cell sources are analyzed. Take home message: It is not possible to engineer 3D thick tissue or organ constructs without effective vascularization. An effective intra-organ vascular system cannot be built by the simple connection of large-diameter vessels and microvessels. Successful engineering of functional human organs suitable for surgical implantation will require concomitant engineering of a ‘built in’ intra-organ branched vascular system. Organ printing enables biofabrication of human organ constructs with a ‘built in’ intra-organ branched vascular tree.

Nanotechnology in vascular tissue engineering: from nanoscaffolding towards rapid vessel biofabrication

The existing methods of biofabrication for vascular tissue engineering are still bioreactor-based, extremely expensive, laborious and time consuming and, furthermore, not automated, which would be essential for an economically successful large-scale commercialization. The advances in nanotechnology can bring additional functionality to vascular scaffolds, optimize internal vascular graft surface and even help to direct the differentiation of stem cells into the vascular cell phenotype. The development of rapid nanotechnology-based methods of vascular tissue biofabrication represents one of most important recent technological breakthroughs in vascular tissue engineering because it dramatically accelerates vascular tissue assembly and, importantly, also eliminates the need for a bioreactor-based scaffold cellularization process.

Changes in the mechanical properties, biochemical contents and wall structure of the human coronary arteries with age and sex.

An experimental study of the mechanical properties, biochemical composition and structure was carried out on the proximal and distal parts of the right and the anterior descending branch of left human coronary arteries. The vessels were removed during an autopsy of 121 males and 84 females being 1 day to 80 years old. The material was divided into six age groups. Branchless segments of vessels 15-20 mm long were cut from proximal and distal parts. The mechanical properties of the coronary arteries were determined by passing fluid at pressures ranging from 0 to 240 mmHg. It was found that the part of the wall of the coronary artery adjacent to the myocardium is thicker in all cases than the other part of the arterial wall. With increasing age the mean thickness of the wall of both coronary arteries increases but the wall-thickening process is non-uniform in nature in both the proximal and distal parts and in the individual layers. The changes in the stretch ratio and tangential modulus in circumferential direction with age and sex were investigated. The greatest changes in the wall thickness and in the mechanical parameters were found for the left coronary artery wall in men over 40 years of age and for the right coronary artery wall in women over 50 years of age. The results of biochemical and densitometric investigations were compared.

Biomechanical and structural properties of the explanted bioprosthetic valve leaflets

Porcine bioprosthesis were treated with 0.625% glutaraldehyde and stabilized under changing pressure from 4 to 30 mmHg. Bovine pericardium and 12 biovalves (of age between 14 days and 80 months) after implantation in the human body were investigated (7 porcine PB and 5 pericardial bioprosthesis--PCB). Circumferential and radial strips from porcine aortic valve leaflets, bovine pericardium and bioprosthetic leaflets were studied in light, transmitting and scanning electron microscopy. Uniaxial load tests were carried out to examine the deformability and strength of these tissues. Microscopic examination of the biovalves revealed that the PB and PCB tissue retained its original architecture, but with alterations in detailed structure. The collagen bundles stuck together with vacuolization between them. There were some areas of the collagen structure fragmentation which could lead to complete necrosis. Eighty months after implantation in patients, the PCB became more extensible and its ultimate strain increases 2.5 times. Ultimate stress decreases in the radial direction from 9.43 to 2.88 MPa, and in the circumferential direction from 9.43 to 6.44 MPa. Forty-eight months after implantation, PB tissues ultimate stress decreases in the circumferential direction from 4.06 to 1.99 MPa. At the same time ultimate strain increases from 13 to 22%. This study is to improve the methods of tissue stabilization in 0.625% glutaraldehyde solution for the first 48 h at cyclic, changing construction of biovalves soft supporting stent after 48 h.

St Jude Epic heart valve bioprostheses versus native human and porcine aortic valves – comparison of mechanical properties

OBJECTIVES The major problem with heart valve bioprostheses made from chemically treated porcine aortic valves is their limited longevity caused by gradual deterioration, which has a causal link with valve tissue mechanical properties. To our best knowledge, there are no published studies on the mechanical properties of modern, commercially available bioprostheses comparing them to native human valves. The objective of this study is to determine the mechanical properties of St Jude Epic bioprostheses and to compare them with native human and porcine aortic valves. METHODS Leaflets from eight porcine aortic valves and six Epic bioprostheses were analyzed using uni-axial tensile tests in radial and circumferential directions. Mechanical properties of human valves have been previously published by our group. Results are represented as mean values+/-S.D. RESULTS Circumferential direction. Modulus of elasticity of Epic bioprostheses in circumferential direction at the level of stress 1.0 MPa is 101.99+/-58.24 MPa, 42.3+/-4.96 MPa for native porcine and 15.34+/-3.84 MPa for human aortic valves. Ultimate stress is highest for Epic bioprostheses 5.77+/-1.94 MPa, human valves have ultimate stress of 1.74+/-0.29 MPa and porcine 1.58+/-0.26 MPa. Ultimate strain in circumferential direction is highest for human valves 18.35+/-7.61% followed by 7.26+/-0.69% for porcine valves and 5.95+/-1.54% for Epic bioprostheses. Radial direction. Modulus of elasticity in radial direction is 9.18+/-1.81 MPa for Epic bioprostheses, 5.33+/-0.61 MPa for native porcine, and 1.98+/-0.15 MPa for human aortic valve leaflets. In the radial direction ultimate stress is highest for Epic bioprostheses 0.7+/-0.21 MPa followed by native porcine valves 0.55+/-0.11 MPa and 0.32+/-0.04 MPa for human valves. For human valves ultimate strain is 23.92+/-4.87%, for native porcine valves 8.57+/-0.8% and 7.92+/-1.74% for Epic bioprostheses. CONCLUSIONS Epic bioprostheses have non-linear stress-strain behavior similar to native valve tissue, but they are significantly stiffer and hence less elastic compared to native porcine and human aortic valves. These differences in mechanical properties may cause variations in stress distribution within leaflets of the bioprosthetic valves and accelerate their deterioration.

Designer ‘blueprint’ for vascular trees: morphology evolution of vascular tissue constructs

Organ printing is a variant of the biomedical application of rapid prototyping technology or layer-by-layer additive biofabrication of 3D tissue and organ constructs using self-assembled tissue spheroids as building blocks. Bioengineering of perfusable intraorgan branched vascular trees incorporated into 3D tissue constructs is essential for the survival of bioprinted thick 3D tissues and organs. In order to design the optimal ‘blueprint’ for digital bioprinting of intraorgan branched vascular trees, the coefficients of tissue retraction associated with post-printing vascular tissue spheroid fusion and remodelling must be determined and incorporated into the original CAD. Using living tissue spheroids assembled into ring-like and tube-like vascular tissue constructs, the coefficient of tissue retraction has been experimentally evaluated. It has been shown that the internal diameter of ring-like and the height of tubular-like tissue constructs are significantly reduced during tissue spheroid fusion. During the tissue fusion process, the individual tissue spheroids also change their shape from ball-like to a conus-like form. A simple formula for the calculation of the necessary number of tissue spheroids for biofabrication of ring-like structures of desirable diameter has been deduced. These data provide sufficient information to design optimal CAD for bioprinted branched vascular trees of desirable final geometry and size.

Biomechanical properties of oesophagus wall under loading

In this investigation, firstly, the biomechanical properties of different parts of oesophagus were determined. Oesophagus stress and strain are the greatest in the cervical part for all age groups. The human oesophagus deforms unevenly, depending on the direction of load in relation to the organs axis, it exhibits anisotropical behaviour. With the age the values of mechanical parameters of the oesophagus wall reduce, in particular beginning from 45 years of age, but the modulus of elasticity increases. Biomechanical properties of the oesophagus depend on the architecture of its structure. By loading the organ in the circumferential direction, microfibrilae rupture and deformation of the muscular fibres occurs. With increase of load, collagenous fibres straighten and microruptures in collagenous fibrilae occur. With stretching of oesophagus longitudinally, collagenous fibres partially preserve their wavy and helical configuration. Therefore, higher resistance of the oesophageal wall occurs in the longitudinal direction.