MSE-BioE+118+2011+TE+1+W - MSE/BioE 118 Lectures#25 Tissue...

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Unformatted text preview: MSE/BioE 118 Lectures #25 Tissue Engineering & Regenerative Medicine 1. 2. 3. 4. 5. Introduction to Tissue Engineering Current Tissue Engineering Products Cells in Tissue Engineering Scaffolds in Tissue Engineering Non-Tissue Engineering Therapeutic Products Replacement Parts §༊ Prosthetic Hips, Knees §༊ Orthopedic Plates, Screws §༊ Dental Implants §༊ Crowns, Fillings §༊ Intraocular Lens §༊ Vascular Grafts §༊ Circulatory Assist Devices §༊ Heart Valves §༊ “Bare” Vascular Stents §༊ Heart Pacemakers §༊ Neurological Stimulators Courtesy Dr. Art Coury Implantable Delivery Devices §༊ Central Venous Catheters §༊ Drug Pumps §༊ Percutaneous Access Devices Gray Area §༊ Drug Eluting Stents §༊ Drug Eluting Pacing Leads §༊ Other Drug-Device Combinations 70-80% of Non-Drug Therapeutic Market Key Components of Tissue Engineering ü༏ Host cells capable of differentiation and organization into tissue specific architectures ü༏ Growth factors and morphogenetic proteins ü༏ Matrix that supports cell attachment, growth, and differentiation A. Mikos, Princ. Tiss. Eng, 2nd Ed., 2000 Whang, K., et al., Polymer, 36, 837, 1995 Langer & Vancanti, Science, 260, 920 (1993) Organogenesis Tissue Engineering New Objectives: Cell, Matrix and Fluid Control Tissue Engineering Functions: §༊ §༊ §༊ §༊ §༊ §༊ §༊ Courtesy Dr. Art Coury Guided Tissue Regeneration Organ Generation/Regeneration Structural Reinforcement Tissue Augmentation/Space Filling Healing Modulation Hypertrophy Inhibition Fouling Prevention Tissue Adhesion/Sealing Adhesion Prevention Enhanced Mitosis/Matrix Deposition Organ Function Modulation Expression of Bio-Actives Tissue Modulation Tissue Ingrowth Integra Depuy Tissue the Clinic Guidance the Abbott 4 mm Col Ethicon Neurosurgery 2006, Tissue Engineering 2011 PLCL Engineered Xenografts & Allografts PGA Cook Biotech Axogen KCI Statistics, World Market Revenues §༊ Pharmaceuticals§༊ Medical Devices§༊ Biotech Products§༊ Combination Products§༊ Cell-Based Products- >$600 Billion (2005) >$200 Billion (2005) >$60 Billion (2005) $5.9 Billion (2004) ~$11.9 Billion (2009 Est.) <<$1 Billion (2006) Courtesy Dr. Art Coury Tissue Engineered Skin Organogenesis - Apligraf ABH - Dermagraft Indication for use: Full thickness venous leg and diabetic foot ulcers Indication for use: Full thickness diabetic foot ulcers Product: Bi-layered. fibroblasts+keratinocytes cultured in bovine collagen matrix. Product: PLGA mesh+newborn foreskin fibroblasts. Supplied - Room temp. (used within 10 days of shipment) Supplied - Cryopreserved INTEGRA® Dermal Regeneration Template Indications for Use: burns and scar contractures Clinical Sequence Implantation of Carticel® Autologous Cultured Chondrocytes Periosteal Flap Defect Biopsy activities of daily living, no sports possible. At 48 months, the mean score was 6.39, good: some limitations with sports but can participate/compensate. The improvement was statistically signiCarticel 7 cant. Table shows the improvement in thecGMP Cell Cincinnati score over time. Overall Modi ed Processing Table 7: Mean Overall Modi ed Cincinnati Score at Baseline and Follow-up Visits Visit Baseline N=154 Month 12 N=146 Month 24 N=131 Month 36 Month 48 N=104 N=101 3.26 5.58 5.92 5.87 6.39 Overall Modi ed Cincinnati Score1 Mean (SD) (1.02) (1.99) (2.08) (2.19) (2.31) 1 Scores are for patients who returned for follow-up. Patients who failed by score criteria are included and patients who failed by surgical criteria are excluded from the scores for timepoints after the failure criteria were met. In addition to the change over time in activity level as measured with the Overall Modi ed Cincinnati Scale, there were similar and consistent changes in knee symptoms and function as measured with the Knee Injury and Osteoarthritis Outcome Score (KOOS), a measure of knee-speci c symptoms and Do Not Refrigerate, Freeze, or Incubate the Carticel Shipping Container or its Contents. Do Not Sterilize. If the Vial is Damaged or Sterility has been Compromised, Do Not Use. 17. PATIENT COUNSELING INFORMATION Patients receiving autologous cultured chondrocytes for treatment of an articular cartilage defect should receive the following information and instructions. • Physical activity should be resumed according to the rehabilitation plan recommended by the physician. Generally, protected weight bearing is recommended for the rst 6 to 8 weeks following implantation. The patient should receive speci c instructions on crutch use, ambulation and weight bearing advancement on the treated limb. Genzyme • If pain starts to develop as the next level of activity is increased, decrease activity to the former level until the pain resolves. • If exercise causes pain and/or swelling, reduce the amount of physical activity. • Swelling should be controlled using ice packs. • Patient adherence to the prescribed rehabilitation program is important and activity at variance from Epicel® Autologous Keratinocyte Graft for Full-Thickness Wounds ® Registered trademark of Genzyme Corporation Surgical results and hospitalization Surgical and hospitalization information mean operative time (hrs) mean EBL (ml) hospitalization (days) spinal level treated L45 (%) L5S1 (%) L56 (%) Investigational Open Surgical Approach 1.6 109.8 3.1 Control Open Surgical Approach 2.0 153.1 3.3 Investigational Laparoscopic Surgical Approach 1.9 146.1 1.2 37 (25.9) 106 (74.1) 0 (0) 32 (23.5) 103 (75.7) 1 (0.7) 21 (15.7) 113 (84.3) 0 (0) Medtronic Sofamor Danek INFUSER While statistically Bone Graft andnotCagesignificant, the operative were and mean bloodthe other two groups Lumbar time lower than for Device Fusion loss for the investigational open surgical approach subjects First Approved Device Using Genetically Engineered required fewer (BMP-2) and the investigational laparoscopic surgical approach subjects Protein hospitalization days. rhBMP-2 in Blood after Intravenous Administration % rhBMP-2 Dose ( ± SD) % rhBMP-2 Dose ( ± SD) X.2.7 Clinical and radiographic effectiveness evaluation at Implant Site following Administration rhBMP-2 of rhBMP-2/ACS Individual subject success was defined as success in each of the primary clinical and radiographic outcome parameters. Success for these100 parameters included: 1. the presence of radiographic fusion; 80 2. an improvement of at least 15 points from the60 baseline Oswestry score; 3. maintenance or improvement in neurological status; 40 4. the presence of no serious adverse event classified as implant-associated or 20 implant/surgical procedure-associated; and 0 5. no additional surgical procedure classified as “Failure.” 10 0 20 30 50 40 30 20 10 0 40 0 10 20 30 40 50 Minutes after Intravenous Injection Days after Implantation Study success was expressed as the number of individual subjects categorized as a success divided by the total number of subjects evaluated. The table below describes the success rates for the individual primary outcome parameters andInteractions Between rhBMP-2 and the Absorbable Collagen Sponge overall success. All IX.2.3.3 success rates were based on the data from the The importance of retaining rhBMP-2 at the implant site for optimal bone formation was 24 month follow- up evaluation and posterior probabilities of success were calculated using Bayesian statistical methods. highlighted in one study in which the heparin-binding domain of rhBMP-2 was removed implantation with ACS, this modified protein was found to leave the implant site much more rapidly than the native protein; for example, only 18% Control Open Surgical Investigational Laparoscopic Approach of the plasmin-cleaved Surgical Approach present at 3 hours, contrasting with 56% for the material was Posterior Mean Posterior Mean native protein. Though the plasmin- cleaved protein was highly active in a cell culture (95% HPD Credible Interval) (95% HPD Credible Interval) assay, 8.1% formation in vivo was substantially reduced, indicating that retention of 8 bone 93.0% (82.6%, 99.3%) (87.9%, 97.5%) bioactive material at the implant site is critical for the desired osteogenic activity. 70.9% 83.0% using plasmin. Following Posterior Probabilities of Success at 24 Months Investigational Open Surgical Approach Primary outcome variable Fusion Oswestry Posterior Mean (95% HPD Credible Interval) 92.8% (88.5%, 96.9%) 71.0% (63.4%, 78.7%) 81.0% (74.5%, 87.9%) 57.1% (49.2%, 65.7%) (63.1%, 79.1%) (75.6%, 90.5%) 81.7% 89.0% As74.9%,local retention of rhBMP-2 is important for localized bone formation, several ( the 88.7%) (83.1%,94.8%) 56.7% 68.0% studies evaluated the effect of potential variables on retention following SC implantation (48.3%, 65.0%) (59.3%, 76.5%) in the rat or orthotopic implantation in the rabbit. The relative retention of rhBMP-2 was Copyright 2004, Medtronic Sofamor Danek FDA Approved July 2, 2002The probability (also called the posterior probability) that the 24 month overall success rhBMP-2 administered (between 0.8 and 2.0 mg/mL) unaffected by the concentration of or to formulation buffer. for amount of rhBMP-2 incorporated into ACS (a measure of rate for the investigational groups was equivalentthethe 24 month success rateThethe the binding of rhBMP-2 to the sponge prior to implantation) had a minimal effect on rhBMP-2 retention in vivo, and no effect on the rate of release of rhBMP-2 into serum in Page 36 of 40 vitro. These results suggest that the release of rhBMP-2 in vivo is independent of the binding of rhBMP-2 to the sponge in vitro, and may be diffusion-controlled. Neurologic Overall success Trade Name® §༊ §༊ §༊ Integra Biobrane Transcyte IX.2.4 Effect of U.S. Tissue Engineering Products Nicotine and Glucocorticosteroids on Implantation with rhBMP2/ACS Type Scaffold Scaffold Scaffold Company Integra Life Sci. Bertek Pharm. ATS, Smith&Neph. Advanced BioHealing For On Market In animal studies, 998sponsor has shown that rhBMP-2/ACS induces bone and fracture 2004-6 1 the repair in the presence of several agents which compromise bone metabolism, for Skin X X example, nicotine and corticosteroids. Skin X X Skin X (X) §༊ §༊ Atrigel Gore-Tex GTR Barrier GTR Barrier QLT, Inc. W.L.Gore Dental Dental X X X X §༊ Osteocel MSCs + Bone Osiris Bone - X §༊ §༊ Carticel Epicel Cultured Cells Cultured Cells Genzyme Genzyme Cartilage Skin X X X X §༊ §༊ Apligraf Dermagraft Cells/Scaffold Cells/Scaffold Skin X X §༊ OrCel Cells/Scaffold Organogenesis ATS, Smith&Neph., Advanced BioHealing Ortec Foot Ulcers Skin X - (X) X §༊ §༊ §༊ §༊ §༊ Regranex Infuse Kepivance OP-1 Putty Gem 21S Growth Factor Growth Factor Growth Factor Growth Factor Growth Factor J&J Medtronic Amgen Stryker Osteohealth Ulcers Spine Mucositis Spine Jaw X - X X X X X Inst. Straumann Jaw - X §༊ Emdogain Adhesion Peptide Courtesy Dr. Art Coury Page 19 of 40 3. Cells in Tissue Engineering •Mesenchymal Stem Cells (adult cells) 3. Cells in Tissue Engineering •Mesenchymal Stem Cells A. Caplan, Methods in Tissue Engineering, Chapter 48 4. Scaffolds in Tissue Engineering Ligand Presentation & Integrin Clustering Material Architecture Material Mechanics Saha, K., et al., Current Opinion in Chemical Biology, 11 (4), 381-387 (2007) 4. Scaffolds in Tissue Engineering • Scaffolds or porous bodies of biodegradable polymers Function • Promote attachment, proliferation, and differentiation of tissue specific cells • Maintain shape and transfer loads • Provide adequate diffusion of nutrients and wastes • Fabricated from biodegradable materials so regenerated tissue is not influenced by residual material Materials • PLA, PGA, PLGA copolymers • Natural hydrogels (collagen, hyaluronic acid, alginate, etc.) • Synthetic hydrogels • Polypeptides Numerous Processing Methods • • Salt leaching Extruded fibers • Emulsion freeze-drying 4. Scaffolds in Tissue Engineering • Synthetic Polymer Hydrogels Chemical Gelation Physical Gelation 4. Scaffolds in Tissue Engineering • Scaffolds or porous bodies of biodegradable polymers Fiber Bonding Salt Leaching Emulsion Freeze-Drying Homogeneous [poly(α-hydroxy acids:PLA, PGA, PLGA] Electrospinning Random'fibers' Zhu et al. Aligned'fibers' Patel&et&al.,&Nano&Le-&(2007)& spinning Apparatus with the Collector of Potential Scaffolds for Skin Tissue Engineering Biomacromolecules, Vol. xxx, No. xx, XXXX F Biomacromolecules, Vol. xxx, No. xx, XXXX linder E Zhu et al. Pharm Res 2011 Figure 6. MTS assay of HDF growth on fibrous mats collected on plate (FM-P), the frame cylinder with rotation speed of 60 rpm (FMHR) and 28 rpm (FM-LR). All values are an estimate of the standard uncertainty expressed as mean ( SD (n ) 4), *p < 0.05. dried at room temperature for 3 d to completely residue prior to further characterization. of Fibrous Mats. The morphologies were obelectron microscope (SEM, Quanta 200, FEI, The ccelerating voltage of 20 kV. The fiber diameter HDF Viability on Fibrous Scaffolds. To monitor cell adhesion and viability on different substrates, the number of cells was determined by using the colorimetric MTS assay. The mechanism behind this assay was that metabolically active cells reacted with tetrazolium salt of the MTS reagent to produce soluble formazan dye that could be observed at 490 nm. The Figure 7. Cross-sections of hematoxylin and eosin stained cell-grown Biomacromolecules 2008 fibrous mats collected on plate (FM-P, A), the frame cylinder with rotation speed of 60 rpm (FM-HR, B) and 28 rpm (FM-LR, C). SEM images of cross-sections of cell-grown fibrous mats of FM-P (D) and FM-LR (E) after a 5 d incubation. Arrows indicate cells residing at the surface and inner part of the scaffold. Organ Printing (Inkjet & SLS) Acta Biomaterialia 2011 Tissue Engineering 2009 Tissue Engineering 2010 Cell Sheets as Biomaterials c orneal reconstruction with autologous oral mucosal epithelium c orneal reconstruction with autologous oral mucosal epithelium A B C D E F c orneal reconstruction with autologous oral mucosal epithelium Figure 2. Transplantation Procedures for Tissue-Engineered Autologous Epithelial-Cell Sheets. Preoperatively, the entire corneal surface was covered by conjunctival tissue with neovascularization (Panel A). In Panel B, conjunctival tissue over the cornea is surgically removed to reexpose transparent corneal stroma. Then, the sheet of tissue-engineered epithelial cells is harvestTable 2. Surgical Outcome in Four Patients Who Received Transplants of Tissue-Engineered Autologous Oral Mucosaled from a temperature-responsive culture insert with the use of a doughnut-shaped supporter ([black-and-white squares] Panel C) and placed Cell Sheets. on the stromal bed (Panel D). The sheet adheres to corneal stroma in a few minutes without sutures, and the supporter is removed (Panel E), leaving the cell sheet on the stroma (Panel F). A video clip can be viewed in the Supplementary Appendix, available with the full text of this article Best Corrected at Patient No. Visual Acuity in Damaged Eye Corneal Opacity (Grade)* Complication Months of Follow-up 1 Month At Last amination with fluorescein sodium staining showed discussion Preoperative† Postoperative Preoperative after Surgery Observation complete reepithelialization of the corneal surface 1 Counting fingers 20/100 2 1 None in all four eyes, revealing the tight junction-mediat- 3Our study shows that tissue-engineered cell sheets 15 2 20/2000 20/25 1 1 None ed barrier function. Corneal transparency was re- 3from autologous oral mucosal epithelium may serve 14 stored without Hand motionof the corneal epitheli- 3as effective substitutes for allografts ofNone tis- 14 any defects limbal 3 20/300 1 1 um. In all eyes, stromal vascularization gradually 3sue in the reconstruction of the corneal and lim- 13 4 20/2000 20/50 1 1 None recurred in the peripheral cornea but not in the cen- bal surfaces. Four patients (four eyes) were contral * The extent vascularization was unlike subepi- masked observers on the basis of the slit-lamp examination with a prezone. This of corneal opacity was graded by three secutively treated with this approach, and corneal thelial vascularization system26 and modifications for ocular-surface diseases. Grade and postoperative trace haze, grade 1 viously described accompanied by conjunctival transparency was restored 0 indicates clear or visual mild since it was 2 moderately dense opacity partially obscuring details of the iris, and grade 3 severely dense opacity ingrowth, opacity, gradelocalized to the deeper stro- acuity improved remarkably (Table 2). During the obscuring show of the intraocular structure. Grading is based period, all corneal surfaces remained ma and did notdetails the abnormally high fluores- follow-up on the opacity observed in all corneal layers, including epithelium, stroma, and endothelium. cein†permeability characteristicwhoconjunctival ep-visual-acuity chart at athere were 0.5 m was assessed by asking whethof could not read a transparent, and distance of no serious complicaThe visual acuity of patients NEJM, 2004 ithelium. could see the number of fingers held up by the examiner. If they could not, visual acuity was assessed by the pations. er they During aability to see hand movement14 months, tient’s mean follow-up period of by the examiner. We developed this strategy on the basis of sevcorneal transparency was maintained (Fig. 3 and eral observations from cell biology and medicine. Table 2). Maximally improved visual acuity was ob- First, in vivo oral mucosal epithelium expresses kertained 6, 2, 10, and 8 weeks after transplantation atin 3, which is also expressed by the corneal epivivo on the substantia propria, which is rich in ves- epithelial cells from various tissue sources, includfor Patients 1 through 4, respectively, and became thelium but not by the epidermis.1,27 Second, the sels. However, the production of antiangiogenic ing the limbus.16 T mucosal tissue from stable thereafter. The length of time until visual excision of a small piece of oralhis method has been clinically factors such as thrombospondin by keratocytes34 applied since the 1980s for the treatment acuity improved seemed to correspond to the length the patient is straightforward, and the resulting of various may limit vascularization to peripheral areas.wound heals within several days without incident skin conditions, including burns and giant nevi, alof time until the corneal stroma became less opaque. We observed that the transplanted cell sheets though the Food and Drug autologous No complications were observed. or scarring. Third, transplantation of Administration classi- became more transparent and achieved smoother, fies these grafts as xenografts. integrated surfaces on the corneal stroma, further In summary, we have shown that sheets of tisn e ngl j 1 resembling normal cornealmed 351;12 september 16, 2004 epithelial cells fabricated ex vivo 193 epithelium; a plateau sue-engineered was reached one to three months after transplanta- from autologous oral mucosal epithelium are effecSMC, CHO, HSF tion. Originally, oral mucosal epithelium, located tive for reconstructing the ocular surface and restorThe New England Journal of Medicine Downloaded fromsubstantia propria, is morphologically distinct ing21, 2011. Forpatients with bilateral total stem-cell on at UC SHARED JOURNAL COLLECTION on November vision in personal use only. No other uses without permission. n e ngl j med 3 51;12 w september 1 6, 2004 1 Medical Society. All rights reserved. Copyright © it is much thicker from corneal epithelium in that2004 Massachusetts191 deficiencies. Long-term follow-up and experience and multilayered and has an irregular surface (Fig. with a large series of patients are needed to assess 1C). The use of temperature-responsive harvesting further the benefits and risks of this method, which The New England Journal of Medicine allows the grafted carrier-free oral mucosal epithe- offers the potential to treat severe ocular diseases Downloaded from at UC SHARED JOURNAL COLLECTION on November 21, 2011. For personal use only. No other uses without permission. lial cells to interact immediately and directly with that are resistant to standard approaches. Copyright © 2004 Massachusetts Medical Society. All rights reserved. patients’ corneal stromal keratocytes without interSupported by Grants-in-Aid for Scientific Research (15390530, ference from cell carriers such as fibrin gel and am- 16200036, and 16300161), the High-Tech Research Center Program, and the Center of Excellence Program for the 21st Century niotic membranes. from the Ministry of Education, Culture, Sports, Science, and TechOur transplantable epithelial-cell sheets used nology in Japan and by the Core Research for Evolution Science and the common 3T3 feeder-layer method originally Technology from the Japan Science and Technology Agency. We are indebted to Professor David Grainger, Colorado State Unideveloped for the production of autologous epider- versity, and Mr. Joseph Yang, Tokyo Women’s Medical University, mal-cell grafts35 and used in the culture of other for their technical review. Cell Aggregates as Biomaterials r eferences 1. Schermer A, Galvin S, Sun TT. Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells. J Cell Biol 1986;103:49-62. 2. Cotsarelis G, Cheng SZ, Dong G, Sun TT, Lavker RM. Existence of slow-cycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: implications on epithelial stem cells. Cell 1989;57:201-9. 3. Thoft RA, Friend J. The X, Y, Z hypothesis of corneal epithelial maintenance. Invest Ophthalmol Vis Sci 1983;24:1442-3. 4. Buck RC. Measurement of centripetal n e ngl j med 3 51;12 w migration of normal corneal epithelial cells in the mouse. Invest Ophthalmol Vis Sci 1985; 26:1296-9. 5. Tseng SC. Concept and application of limbal stem cells. Eye 1989;3:141-57. 6. Nishida K. Tissue engineering of the cornea. Cornea 2003;22:Suppl 7:S28-S34. 1195 september 1 6, 2004 The New England Journal of Medicine Downloaded from at UC SHARED JOURNAL COLLECTION on November 21, 2011. For personal use only. No other uses without permission. Copyright © 2004 Massachusetts Medical Society. All rights reserved. Biomaterials 2009 Vascular TE and Tissue Histology Nature Medicine 2006 ...
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