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1.
Biomaterials ; 35(29): 8297-304, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24997479

ABSTRACT

Biologic scaffold materials are used for repair and reconstruction of injured or missing tissues. Such materials are often composed of allogeneic or xenogeneic extracellular matrix (ECM) manufactured by decellularization of source tissue, such as dermis. Dermal ECM (D-ECM) has been observed to degrade and remodel in vivo more slowly than other biologic scaffold materials, such as small intestinal submucosa (SIS-ECM). Histologic examination is a common method for evaluating material degradation, but it lacks sensitivity and is subject to observer bias. Utilization of (14)C-proline labeled ECM is a quantitative alternative for measuring degradation of ECM scaffolds. Using both methods, the amount of degradation of D-ECM and SIS-ECM was determined at 2, 4, and 24 weeks post-implantation in a rodent model. Results utilizing (14)C liquid scintillation counting (LSC) analysis showed distinct differences in degradation at the three time points. D-ECM material in situ stayed the same at 76% remaining from 2 to 4 weeks post-implantation, and then decreased to 44% remaining at 24 weeks. In the same time period, implanted SIS-ECM material decreased from 72% to 13% to 0%. Visual examination of device degradation by histology overestimated degradation at 2 weeks and underestimated device degradation at 24 weeks, compared to the (14)C method.


Subject(s)
Biocompatible Materials/metabolism , Dermis/metabolism , Extracellular Matrix/metabolism , Tissue Scaffolds/chemistry , Animals , Biocompatible Materials/chemistry , Carbon Radioisotopes/analysis , Carbon Radioisotopes/metabolism , Dermis/chemistry , Dermis/ultrastructure , Extracellular Matrix/chemistry , Extracellular Matrix/ultrastructure , Female , Rats, Sprague-Dawley , Scintillation Counting/methods , Swine
2.
Biomaterials ; 35(25): 6838-49, 2014 Aug.
Article in English | MEDLINE | ID: mdl-24856104

ABSTRACT

The host response to implanted biomaterials is a highly regulated process that influences device functionality and clinical outcome. Non-degradable biomaterials, such as knitted polypropylene mesh, frequently elicit a chronic foreign body reaction with resultant fibrosis. Previous studies have shown that an extracellular matrix (ECM) hydrogel coating of polypropylene mesh reduces the intensity of the foreign body reaction, though the mode of action is unknown. Macrophage participation plays a key role in the development of the foreign body reaction to biomaterials, and therefore the present study investigated macrophage polarization following mesh implantation. Spatiotemporal analysis of macrophage polarization was conducted in response to uncoated polypropylene mesh and mesh coated with hydrated and dry forms of ECM hydrogels derived from either dermis or urinary bladder. Pro-inflammatory M1 macrophages (CD86+/CD68+), alternatively activated M2 macrophages (CD206+/CD68+), and foreign body giant cells were quantified between 3 and 35 days. Uncoated polypropylene mesh elicited a dominant M1 response at the mesh fiber surface, which was decreased by each ECM coating type beginning at 7 days. The diminished M1 response was accompanied by a reduction in the number of foreign body giant cells at 14 and 35 days, though there was a minimal effect upon the number of M2 macrophages at any time. These results show that ECM coatings attenuate the M1 macrophage response and increase the M2/M1 ratio to polypropylene mesh in vivo.


Subject(s)
Extracellular Matrix/chemistry , Macrophages/chemistry , Polypropylenes/chemistry , Surgical Mesh , Animals , Coated Materials, Biocompatible/chemistry , Female , Fibrosis/pathology , Foreign-Body Reaction , Hydrogels/chemistry , Immunity, Innate/drug effects , Materials Testing , Prostheses and Implants , Rats , Rats, Sprague-Dawley
3.
J Biomed Mater Res A ; 102(1): 234-46, 2014 Jan.
Article in English | MEDLINE | ID: mdl-23873846

ABSTRACT

Surgical mesh devices composed of synthetic materials are commonly used for ventral hernia repair. These materials provide robust mechanical strength and are quickly incorporated into host tissue; factors that contribute to reduced hernia recurrence rates. However, such mesh devices cause a foreign body response with the associated complications of fibrosis and patient discomfort. In contrast, surgical mesh devices composed of naturally occurring extracellular matrix (ECM) are associated with constructive tissue remodeling, but lack the mechanical strength of synthetic materials. A method for applying a porcine dermal ECM hydrogel coating to a polypropylene mesh is described herein with the associated effects upon the host tissue response and biaxial mechanical behavior. Uncoated and ECM coated heavy-weight BARD™ Mesh were compared to the light-weight ULTRAPRO™ and BARD™ Soft Mesh devices in a rat partial thickness abdominal defect overlay model. The ECM coated mesh attenuated the pro-inflammatory response compared to all other devices, with a reduced cell accumulation and fewer foreign body giant cells. The ECM coating degraded by 35 days, and was replaced with loose connective tissue compared to the dense collagenous tissue associated with the uncoated polypropylene mesh device. Biaxial mechanical characterization showed that all of the mesh devices were of similar isotropic stiffness. Upon explanation, the light-weight mesh devices were more compliant than the coated or uncoated heavy-weight devices. This study shows that an ECM coating alters the default host response to a polypropylene mesh, but not the mechanical properties in an acute in vivo abdominal repair model.


Subject(s)
Coated Materials, Biocompatible/chemistry , Extracellular Matrix/chemistry , Foreign-Body Reaction/metabolism , Foreign-Body Reaction/pathology , Materials Testing , Polypropylenes/chemistry , Animals , Female , Rats , Rats, Sprague-Dawley , Surgical Mesh
4.
Biomaterials ; 28(26): 3786-96, 2007 Sep.
Article in English | MEDLINE | ID: mdl-17574665

ABSTRACT

Calcium phosphate cement (CPC) is highly promising for clinical uses due to its in situ-setting ability, excellent osteoconductivity and bone-replacement capability. However, the low strength limits its use to non-load-bearing applications. The objectives of this study were to develop a layered CPC structure by combining a macroporous CPC layer with a strong CPC layer, and to investigate the effects of porosity and layer thickness ratios. The rationale was for the macroporous layer to accept tissue ingrowth, while the fiber-reinforced strong layer would provide the needed early-strength. A biopolymer chitosan was incorporated to strengthen both layers. Flexural strength, S (mean+/-sd; n=6) of CPC-scaffold decreased from (9.7+/-1.2) to (1.8+/-0.3) MPa (p<0.05), when the porosity increased from 44.6% to 66.2%. However, with a strong-layer reinforcement, S increased to (25.2+/-6.7) and (10.0+/-1.4) MPa, respectively, at these two porosities. These strengths matched/exceeded the reported strengths of sintered porous hydroxyapatite implants and cancellous bone. Relationships were established between S and the ratio of strong layer thickness/specimen thickness, a/h:S=(17.6 a/h+3.2) MPa. The scaffold contained macropores with a macropore length (mean+/-sd; n=147) of (183+/-73) microm, suitable for cell infiltration and tissue ingrowth. Nano-sized hydroxyapatite crystals were observed to form the scaffold matrix of CPC with chitosan. In summary, a layered CPC implant, combining a macroporous CPC with a strong CPC, was developed. Mechanical strength and macroporosity are conflicting requirements. However, the novel functionally graded CPC enabled a relatively high strength and macroporosity to be simultaneously achieved. Such an in situ-hardening nano-apatite may be useful in moderate stress-bearing applications, with macroporosity to enhance tissue ingrowth and implant resorption.


Subject(s)
Bone Cements/chemistry , Calcium Phosphates/chemistry , Chitosan/chemistry , Models, Chemical , Nanostructures/chemistry , Adhesiveness , Compressive Strength , Computer Simulation , Crystallization/methods , Elasticity , Hardness , Materials Testing , Nanostructures/ultrastructure , Particle Size , Porosity , Tensile Strength
5.
J Mater Sci Mater Med ; 18(7): 1345-53, 2007 Jul.
Article in English | MEDLINE | ID: mdl-17277972

ABSTRACT

Calcium phosphate cement (CPC) sets in situ to form resorbable hydroxyapatite and is promising for orthopaedic applications. However, it requires on-site powder-liquid mixing during surgery, which prolongs surgical time and raises concerns of inhomogeneous mixing. The objective of this study was to develop a premixed CPC scaffold with macropores suitable for tissue ingrowth. To avoid the on-site powder-liquid mixing, the CPC paste was mixed in advance and did not set in storage; it set only after placement in a physiological solution. Using 30% and 40% mass fractions of mannitol porogen, the premixed CPC scaffold with fibers had flexural strength (mean +/- sd; n = 5) of (3.9 +/- 1.4) MPa and (1.8 +/- 0.8) MPa, respectively. The scaffold porosity reached (68.6 +/- 0.7)% and (74.7 +/- 1.2)%, respectively. Osteoblast cells colonized in the surface macropores of the scaffold and attached to the hydroxyapatite crystals. Cell viability values for the premixed CPC scaffold was not significantly different from that of a conventional non-premixed CPC known to be biocompatible (P > 0.1). In conclusion, using fast-dissolving porogen and slow-dissolving fibers, a premixed macroporous CPC scaffold was developed with strength approaching the reported strengths of sintered porous hydroxyapatite implants and cancellous bone, and non-cytotoxicity similar to a biocompatible non-premixed CPC.


Subject(s)
Bone Cements/chemistry , Bone Cements/pharmacology , Calcium Phosphates/administration & dosage , Calcium Phosphates/chemistry , Cell Survival/drug effects , 3T3 Cells , Adhesiveness , Animals , Compressive Strength , Elasticity , Hardness , Materials Testing , Mice , Porosity , Stress, Mechanical
6.
Dent Mater ; 23(4): 433-41, 2007 Apr.
Article in English | MEDLINE | ID: mdl-16678895

ABSTRACT

OBJECTIVES: Calcium phosphate cement (CPC) is a promising material for dental, periodontal, and craniofacial repairs. However, its use requires on-site powder-liquid mixing that increases the surgical placement time and raises concerns of insufficient and inhomogeneous mixing. The objective of this study was to determine a formulation of premixed CPC (PCPC) with rapid setting, high strength, and good in vitro cell viability. METHODS: PCPCs were formulated from CPC powder+non-aqueous liquid+gelling agent+hardening accelerator. Five PCPCs were thus developed: PCPC-Tartaric, PCPC-Malonic, PCPC-Citric, PCPC-Glycolic, and PCPC-Malic. Formulations and controls were compared for setting time, diametral tensile strength, and osteoblast cell compatibility. RESULTS: Setting time (mean+/-S.D.; n=4) for PCPC-Tartaric was 8.2+/-0.8 min, significantly less than the 61.7+/-1.5 min for the Premixed Control developed previously (p<0.001). On 7th day immersion, the diametral tensile strength of PCPC-Tartaric reached 6.5+/-0.8 MPa, higher than 4.5+/-0.8 MPa of Premixed Control (p=0.036). Osteoblast cells displayed a polygonal morphology and attached to the nano-hydroxyapatite crystals in the PCPCs. All cements had similar live cell density values (p=0.126), indicating that the new PCPCs were as cell compatible as a non-premixed CPC control known to be biocompatible. Each of the new PCPCs had a cell viability that was not significantly different (p>0.1) from that of the non-premixed CPC control. SIGNIFICANCE: PCPCs will eliminate the powder-liquid mixing during surgery and may also improve the cement performance. The new PCPCs supported cell attachment and yielded a high cell density and viability. Their mechanical strengths approached the reported strengths of sintered porous hydroxyapatite implants and cancellous bone. These nano-crystalline hydroxyapatite cements may be useful in dental, periodontal, and craniofacial repairs.


Subject(s)
Calcium Phosphates/toxicity , Dental Cements/toxicity , Hydroxyapatites/toxicity , Osteoblasts/drug effects , 3T3 Cells , Animals , Cell Adhesion , Cell Survival , Dental Cements/chemical synthesis , Dental Stress Analysis , Durapatite , Hydroxyapatites/chemical synthesis , Mice , Nanostructures , Tartrates/chemistry , Tensile Strength
7.
Biomaterials ; 26(24): 5002-14, 2005 Aug.
Article in English | MEDLINE | ID: mdl-15769536

ABSTRACT

Although calcium phosphate cement (CPC) is promising for bone repair, its clinical use requires on site powder-liquid mixing. To shorten surgical time and improve graft properties, it is desirable to develop premixed CPC in which the paste remains stable during storage and hardens only after placement into the defect. The objective of this study was to develop premixed CPC with rapid setting when immersed in a physiological solution. Premixed CPCs were formulated using the following approach: Premixed CPC = CPC powder + nonaqueous liquid + gelling agent + hardening accelerator. Three premixed CPCs were developed: CPC-monocalcium phosphate monohydrate (MCPM), CPC-chitosan, and CPC-tartaric. Setting time for these new premixed CPCs ranged from 5.3 to 7.9 min, significantly faster than 61.7 min for a premixed control CPC reported previously (p < 0.05). SEM revealed the formation of nano-sized needle-like hydroxyapatite crystals after 1 d immersion and crystal growth after 7 d. Diametral tensile strength for premixed CPCs at 7 d ranged from 2.8 to 6.4 MPa, comparable to reported strengths for cancellous bone and sintered porous hydroxyapatite implants. Osteoblast cells attained a normal polygonal morphology on CPC-MCPM and CPC-chitosan with cytoplasmic extensions adhering to the nano-hydroxyapatite crystals. In summary, fast-setting premixed CPCs were developed to avoid the powder-liquid mixing in surgery. The pastes hardened rapidly once immersed in physiological solution and formed hydroxyapatite. The cements had strengths matching those of cancellous bone and sintered porous hydroxyapatite and non-cytotoxicity similar to conventional non-premixed CPC.


Subject(s)
Biocompatible Materials/adverse effects , Biocompatible Materials/chemistry , Bone Cements/chemistry , Calcium Phosphates/adverse effects , Calcium Phosphates/chemistry , Cell Survival/drug effects , 3T3 Cells , Adhesiveness , Animals , Biocompatible Materials/analysis , Biocompatible Materials/therapeutic use , Bone Cements/adverse effects , Bone Cements/analysis , Bone Cements/therapeutic use , Calcium Phosphates/analysis , Calcium Phosphates/therapeutic use , Cementation/methods , Elasticity , Fractures, Bone/drug therapy , Hardness , Humans , Manufactured Materials/analysis , Materials Testing , Mice , Tensile Strength
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