Innovation, competitiveness, reliability.
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1 ACTIGEN
2 Innovation, competitiveness, reliability.
3 UBGEN : Generate new business by regenerating health. To generate is to give life, get results, profitability, news. To regenerate is to take care of, rebuild, heal. UBGEN s name, which stands for Ultra Bone Generation, contains the genes of origin of new innovative business models in the biotechnology industry for the repair and regeneration of hard and soft tissue. Marco Manno, founder and CEO explains. The simple way: Intuition for business. Some companies are inherited, others are acquired; others are born as a brain flash, running through the mind of their founder. At the International Dental Show in Cologne, in March 2007, as I searched in vain for a manufacturer of biomaterials to distribute on the Italian market, the thought occurred to me to create a start-up company dedicated to the circulation and commercialisation of innovative biomedical technology for tissue engineering in the dental industry. The opportunity to go forward with my insight came during the Masters at Sole 24Ore, in which my project work was awarded as the best among the 7 presented by the faculty. The pride and happiness made me realise that the dream could well become reality, it was up to me. 3
4 Our products: Concreteness and results.
5 The courageous way: The practice of innovative models. Thanks to the support and valuable guidance of fellow entrepreneurs and professionals, the idea was transformed into a studio project, which also involved the Department of Biology at the University of Padua. The morning that the Rector of the University at the time, Prof. Rizzuto, confirmed the willingness of his team to work on the development of the bone matrix de-antigenation process, and it was beginning to get serious: I was sitting at the table in his office, in full suit and tie, outwardly calm and collected, while the excitement of reaching my goal made my heart pound. The die was cast. 5
6 The complex way: Perseverance through difficulties. Once the bovine bone de-proteinisation process was defined, we moved on to the animal testing stage, the birth of the UBGEN start-up, to be equipped with a clean-room and with all machinery required to obtain the final product, testing on human beings, right up to the great unknown: Obtaining certifications from the National Institute of Health, which required compliance with innumerable formalities and appeared to be one of the most difficult issues to be addressed for the success of the project. The bureaucracy didn t help our work: Each time we solved a problem, another one would pop up, then another, in an interminable, depressing sequence that seemed never-ending, but the tenacity, perseverance and constancy of my wife and all the staff led us to the result: While we were fighting with the paperwork and technical dossiers required for certifications, we didn t know yet that we'd be the first to obtain them at national level. 6
7 The conscious way: The power of decision. Every idea requires great courage to achieve it. When it was time to start, I really found myself alone: People who were initially so enthusiastic drew back, but the clarity of my goals, and the support of my wife and the staff gave me strength and determination. Conscious of my role to generate business in a world that changes very quickly, I have been committed from the outset to looking beyond and diversifying products and services, while maintaining focus on the chosen field: Tissue engineering. 7
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9 ACTIGEN The new and exclusive coating of the implant surface in collagen. A biological surface to promote implant osseointegration.
10 THE STRENGTH OF COMPATIBILITY The implant coating to date Today s most widely used dental implants come with two types of surface coatings: DAE (Double Acid-Etched) and SLA (Sandblasted and Acid- Etched). For this to happen, cells must be able perceive surface roughness as compatible with their own size. 1-2 DAE (Double Acid-Etched) SLA (Sandblasted and Acid-Etched) 1. Boyan BD et al; Titanium surface roughness alters responsiveness of MG63 osteoblast-like cells to 1 alpha, 25-(OH)2D3; J Biomed Mater Res Jan;39(1): Lincks J et al: Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition; Biomaterials 1998; 19(23):
11 Osseointegration and implant stability The concepts and ideas behind SLA and DAE surfaces have enabled significant progress in the design of implant fixtures. However, they have not yet solved the problem of immediate post-extractive implant placement in fresh sockets, which is still associated with poor primary stability. Osseointegration is indeed favoured by primary mechanical stability, later reinforced by biological stability (graph 1). GRAPH 1 Stability of dental implants over time 100 Total stability Stability % Biological stability Mechanical stability 0 Time 11
12 UBGEN process analysis and research Starting from today s implant coating technologies, UBGEN has invested in the search for a biological surface that accelerates and increases the osseointegration of implant fixtures. The chemical composition and topography of surfaces are used as a tool to influence cell behaviour, and therefore also the bone regeneration process. For some time now, it has been suggested that implant surfaces may be activated through functionalisation with a number of biologically active molecules for bone formation: Including synthetic peptides, growth factors, and others. 3-4 UBGEN followed this direction for the development of Actigen : An exclusive implant surface coating. 3. Morra M. Biochemical modification of titanium surfaces: Peptides and ECM proteins. Eur Cell Mater Jul 24;12: De Jonge LT et al. Organic-inorganic surface modifications for titanium implant surfaces. Pharm Res Oct;25(10): doi: /s Epub 2008 May
13 Actigen : The evolution of implant coating A significant result in terms of osseointegration was achieved by immobilising dermal collagen (type I) over etched surfaces, since it is the same type of collagen that is the major organic component of bone and supports its vital processes. 5-7 During the regeneration processes, osteoblast cells initially deposit a collagen matrix, which is then mineralised. This collagen matrix exerts a number of positive biological effects, including the adhesion of osteoblasts and osteoclasts, it is a cofactor for several growth factors and many other mechanisms For UBGEN, the use of collagen was a choice that proved crucial to the creation of a biological solution that anticipates future needs in the field of dentistry. Collagen-based implant surface coating µm Collagen surface - 10 µm 5. Morra M et al. Surface engineering of titanium by collagen immobilization. Surface characterization and in vitro and in vivo studies. Biomaterials Nov;24(25): Morra M et al. Collagen I-coated titanium surfaces: mesenchymal cell adhesion and in vivo evaluation in trabecular bone implants. J Biomed Mater Res A Sep 1;78(3): Regazzoni C. et al. Type I collagen induces expression of bone morphogenetic protein receptor type II Biochem. Biophys. Res. Commun. 2001; 4 ; 283(2):
14 The functions of collagen Stimulation mechanisms that allow the physiological development of bone regeneration. Angiogenesis Collagen carries out chemotactic action on monocytes/macrophages cell lines, from which osteoclasts derive. In turn, these cells promote angiogenesis by secreting angiogenic growth factors. Collagen can also activate the receptors on the cell membrane of platelets, facilitating hemostasis. Remodelling Collagen carries out chemotactic action on osteoclast precursors; these cells collaborate with osteoblasts in bone rearranging and remodelling through their action on mineral component resorption. Osteoblastic activity When collagen binds to fibronectin, it promotes the anchorage of mesenchymal stem progenitors, on which it exerts its chemotactic action and induces osteoblast differentiation. Osteoconduction and guided bone regeneration Collagen can both increase the osteoblast proliferation rate and also guide the regeneration of connective tissue. 14
15 The presence of ACTIGEN on the implant surface therefore causes the first stage of new bone formation, providing a matrix that is ready for mineralisation by the relevant cells (osteoblasts) and for subsequent bone growth, while also promoting osseointegration with biochemical stimulation. In vitro tests To confirm the properties of collagen in cell adhesion and cell differentiation stimulation, some in vitro tests were published on Actigen -coated implants. Tests on the production of the enzyme alkaline phosphatase (ALP) by osteoblasts cultured on titanium bars and titanium bars coated with collagen. The results after 3 and 7 days of culture show that the collagen-coated titanium produced more ALP, the main marker of osteoblastic activity, confirming the role of collagen inducing pro-osteogenic activity. GRAPH 2 3 days Standard collagen 3 days Actigen 7 days Standard collagen 7 days Actigen ALP activity μm p-nitropheno/mg protein/minute 15
16 Tests on cell growth of human mesenchymal cells (HMC) adhering to a titanium bar and titanium bar coated with collagen. After twenty-four hours of culture, the cell nuclei were stained with a fluorophore, thus highlighting a greater cell density on the titanium bar coated with collagen. Observing the same cultures under an electron microscope highlighted a broader and less polarized cell shape, indicating a cuboidal phenotype of the osteogenic differentiation process. The collagen coating was applied to titanium screws which were then inserted into rabbit femurs for an in vivo assessment after four weeks. The trabecular bone showed improvements of the bone-implant contact surface compared to the control (non-treated titanium implant) which corresponds to faster regeneration of the bone surrounding the implant site. A second set of implants were inserted in rabbit femurs and a histomorphometric analysis was performed at two and four weeks. These analyses showed a significant increase of the bone-implant contact surface at two weeks, while the bone healing was complete on both implant surfaces at four weeks. This information supports the hypothesis that collagen induces faster bone production at the interface with the implant in accordance with the biological role of collagen. GRAPH 3 Histomorphometric assessment of Ti and ColTi implants Ti Ti ColTi ColTi %BIC 16
17 Case report A case report carried out on animals in 2016 included 160 Actigen -coated implants following a strict surgical protocol and clinical follow-up, in order to demonstrate the effectiveness of the coating with the following guidelines: _ Placement in native bone D4; _ Uncovering after 10 weeks; _ ASA 1 patient. Two tests were carried out at the uncovering to demonstrate the attainment of implant stability: The percussion test and the torque removal test at 20 N cm. All surgical steps were documented with X-rays and photographs. After one year, all implants were correctly osseointegrated after the uncovering of the implant at ten weeks. 17
18 Collagen surface - 1µm ACTIGEN : The difference lies in the result The analysis of histomorphometry data on ACTIGEN -coated implants has shown that the localisation of collagen molecules at the interface has increased both the percentage of bone-implant contact and bone growth within the implant threads in a statistically significant way. 6 Results show that the ACTIGEN surface allows an increase of up to 3 times in BIC (Bone Interface Contact) in only two weeks, compared to a traditional surface. Why Collagen? _ Stimulates angiogenesis; _ Promotes cell adhesion; _ Promotes osteoblastic behaviour; _ Promotes remodelling and mineralisation. What benefits does it provide? _ Prepares the implant surface to be rapidly colonised by bone tissue; _ Increases BIC; _ Allows for faster bone growth; _ Increases bone quantity and quality; _ Stimulates platelet activation; _ It is a Class III certified device, complying with strict requirements to ensure patient safety and protection. 6. Morra M et al. Surface analysis and effects on interfacial bone microhardness of collagen-coated titanium implants: a rabbit model. Int J Oral Maxillofac Implants Jan-Feb;20(1):
19 GRAPH 4 Bone Implant Contact DAE - SLA / BIC = 20% DAE - SLA / BIC = 20% Actigen / BIC = 60% Actigen / BIC = 60%
20 BIBLIOGRAPHY 1. Boyan BD et al; Titanium surface roughness alters responsiveness of MG63 osteoblast-like cells to 1 alpha, 25-(OH)2D3; J Biomed Mater Res Jan;39(1): Lincks J et al: Response of MG63 osteoblast-like cells to titanium and titanium alloy is dependent on surface roughness and composition; Biomaterials 1998; 19(23): Morra M. Biochemical modification of titanium surfaces: Peptides and ECM proteins. Eur Cell Mater Jul 24;12: De Jonge LT et al. Organic-inorganic surface modifications for titanium implant surfaces. Pharm Res Oct;25(10): doi: /s Epub 2008 May Morra M et al. Surface engineering of titanium by collagen immobilization. Surface characterization and in vitro and in vivo studies. Biomaterials Nov;24(25): Morra M et al. Surface analysis and effects on interfacial bone microhardness of collagen-coated titanium implants: a rabbit model. Int J Oral Maxillofac Implants Jan-Feb;20(1): Morra M et al. Collagen I-coated titanium surfaces: mesenchymal cell adhesion and in vivo evaluation in trabecular bone implants. J Biomed Mater Res A Sep 1;78(3): Baslé MF et al. Type I collagen in xenogenic bone materials regulates attachement and spreading of osteoblasts over the beta 1 integrin subunit. Orthopade 1998; 27(2): Paralkar VM et al. Interaction of osteogenin, a heparin binding bone morphogenetic protein, with type IV collagen. J Biol Chem Oct 5;265(28): Toroian D et al. The size exclusion characteristics of type I collagen: implication for the role of noncollagenous bone constituents in mineralization. J. Biol. Chem. 1990; 5; 265(28): Sampath T.K. Dissociative extraction and recostruction of extracellular matrix components. 12. Green J et al. Cell-matrix interaction in bone: type I collagen modulates signal transduction in osteoblast like cells. Am. J. Physiol. 1995; 268(5 Pt 1): C Liu G. et al. Effect of type I collagen on the adhesion, proliferation and osteoblastic gene expression of bone marrow-derived mesenchymal stem cells. Chin J. Traumatol. 2004; 7(6): Mizuno M. et al. Type I collagen-induced osteoblastic differentiation of bone-marrow cells mediated by collagen-alpha2beta1 integrin interaction. J. Cell Physiol. 2000; 184(2): Gungormus M. et al.the effect of osteogenesis of type I collagen applied to experimental bone defects. Dent. Traumatol. 2004; 7(6). 16. Regazzoni C. et al. Type I collagen induces expression of bone morphogenetic protein receptor type II Biochem. Biophys. Res. Commun. 2001; 4 ; 283(2): Morra M, Cassinelli C, Cascardo G, Mazzucco L, Borzini P, Fini M, Giavaresi G, Giardino R. Collagen I-coated titanium surfaces: mesenchymal cell adhesion and in vivo evaluation in trabecular bone implants. J Biomed Mater Res A 2006; 78(3): Morra M, Cassinelli C, Cascardo G, Bollati D, Baena RR. Gene expression of markers of osteogenic differentiation of human mesenchymal cells on collagen I-modified microrough titanium surfaces. J Biomed Mater Res A 2011; 96(2): Morra M, Cassinelli C, Cascardo G, Bollati D, Rodriguez y Baena R. Multifunctional implant surfaces: Surface characterization and bone response to acid-etched Ti Implants surface modified by fibrillar collagen I. J Biomed Mater Res A 2010;94(1):
21 UBGEN: OUR PRODUCTS RE-BONE A specific line of bone substitutes to promote the regeneration of tissues in bone and reconstructive surgery. Available in versions: _ Block _ Cancellous and cortico-cancellous granules _ Syringe SHELTER membrane A specific line of cell separators to promote the regeneration of tissues in bone and reconstructive surgery. Available in versions: _ Slow absorption pericardium membrane _ Fast absorption pericardium membrane Multipurpose GF-ONE Centrifuge for applications with medical devices. A counter-top device specifically designed for separating blood components. It is managed by a microprocessor which allows users to set speed and centrifugation time, with the ability to customise programmes. ACTIGEN A new and exclusive collagen-based implant surface coating. A biological surface to promote osseointegration. Bone & Tissue Management Surgical instruments and PTFE sutures to support the most advanced oral surgery techniques. 21
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24 UBGEN srl Viale del Lavoro, Vigonza (PD) Italy T info@ubgen.com ubgen.com
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