Composite adhesives composed of biodegradable scaffolds impregnated with a biological or synthetic adhesive were investigated for use in wound closure as an alternative to using either one of the adhesives alone. Two different scaffold materials were investigated: (i) a synthetic biodegradable material fabricated from poly(L-lactic-co-glycolic acid); and (ii) a biological material, small intestinal sub mucosa, manufactured by Cook BioTech. The biological adhesive was composed of 50%(w/v) bovine serum albumin solder and 0.5mg/ml indocyanine green dye mixed in deionized water, and activated with an 808-nm diode laser. The synthetic adhesive was Ethicon's Dermabond, a 2-octyl-cyanoacrylate. The tensile strength of skin incisions repaired ex vivo in a rat model, by adhesive alone or in combination with a scaffold, as well as the time-to-failure, were measured and compared. The tensile strength of repairs formed using the scaffold-enhanced biological adhesives were on average, 80% stronger than their non-enhanced counterparts, with an accompanying increase in the time-to-failure of the repairs. These results support the theory that a scaffold material with an irregular surface that bridges the wound provides a stronger, more durable and consistent adhesion, due to the distribution of the tensile stress forces over the many micro-adhesions provided by the irregular surface, rather than the one large continuous adhesive contact. This theory is also supported by several previous ex vivo experiments demonstrating enhanced tensile strength of irregular versus smooth scaffold surfaces in identical tissue repairs performed on bovine thoracic aorta, liver, spleen, small intestine and lung tissue.
Our Scaffold-Enhanced Biological Adhesive (SEBA) system was investigated as an alternative to sutures or adhesives alone for repair of wounds. Two scaffold materials were investigated: (i) a synthetic biodegradable material fabricated from poly(L-lactic-co-glycolic acid); and (ii) a biologic material, small intestinal submucosa, manufactured by Cook BioTech. Two adhesive materials were also investigated: (i) a biologic adhesive composed of 50%(w/v) bovine serum albumin solder and 0.5mg/ml indocyanine green dye mixed in deionized water, and activated with an 808-nm diode laser; and (ii) Ethicon’s Dermabond, a 2-octyl-cyanoacrylate. The tensile strength and time-to-failure of skin incisions repaired in vivo in a rat model were measured at seven days postoperative. Incisions closed by protein solder alone, by Dermabond alone, or by suture, were also tested for comparison. The tensile strength of repairs formed using the SEBA system were 50% to 65% stronger than repairs formed by suture or either adhesive alone, with significantly less variations within each experimental group (average standard deviations of 15% for SEBA versus 38% for suture and 28% for adhesive alone). In addition, the time-to-failure curves showed a longevity not previously seen with the suture or adhesive alone techniques. The SEBA system acts to keep the dermis in tight apposition during the critical early phase of wound healing when tissue gaps are bridged by scar and granulation tissue. It has the property of being more flexible than either of the adhesives alone and may allow the apposed edges to move in conjunction with each other as a unit for a longer period of time and over a greater range of stresses than adhesives alone. This permits more rapid healing and establishment of integrity since the microgaps between the dermis edges are significantly reduced. By the time the scaffolds are sloughed from the wound site, there is greater strength and healing than that produced by adhesive alone or by wounds following suture removal. This hypothesis is supported by the data of this study, as well as, the acute tensile strength data of Part I of this study.
An ex vivo study was conducted to determine the effect of the irregularity of the scaffold surface on the tensile strength of repairs formed using our Scaffold-Enhanced Biological Adhesive (SEBA). Two different scaffold materials were investigated: (i) a synthetic biodegradable material fabricated from poly(L-lactic-co-glycolic acid); and (ii) a biological material, small intestinal submucosa, manufactured by Cook BioTech. The scaffolds were doped with protein solder composed of 50%(w/v) bovine serum albumin solder and 0.5mg/ml indocyanine green dye mixed in deionized water, and activated with an 808-nm diode laser. The tensile strength of repairs performed on bovine thoracic aorta, liver, spleen, small intestine and lung, using the smooth and irregular surfaces of the above scaffold-enhanced materials were measured and the time-to-failure was recorded. The tensile strength of repairs formed using the irregular surfaces of the scaffolds were consistently higher than those formed using the smooth surfaces of the scaffolds. The largest difference was observed on repairs formed on the aorta and small intestine, where the repairs were, on average, 50% stronger using the irregular versus the smooth scaffold surfaces. In addition, the time-to-failure of repairs formed using the irregular surfaces of the scaffolds were between 50% and 100% longer than that achieved using the smooth surfaces of the scaffolds. It has previously been shown that distributing or dispersing the adhesive forces over the increased surface area of the scaffold, either smooth or irregular, produces stronger repairs than albumin solder alone. The increase in the absolute strength and longevity of repairs seen in this new study when the irregular surfaces of the scaffolds are used is thought to be due to the distribution of forces between the many independent micro-adhesions provided by the irregular surfaces.
The ability to reproduce strong repairs is essential to establishing the reliability of laser-tissue soldering techniques and advancing their use to the clinical setting. While some thermal damage is necessary to achieve a viable solder-tissue bond, excessive thermal damage leads to decreased flexibility and strength of the repair. In addition, if the temperature at the solder/tissue interface is too low, inadequate solder-tissue bonding will occur to provide a strong repair. This suggests the presence of an optimal temperature for laser-tissue repair. The choice of solder material presents another challenge to the reproducibility of strong repairs. The emerging use of chromophore-enhanced solder-doped polymer scaffolds offers numerous advantages over more traditional liquid and solid solders composed of serum albumin and an absorbing chromophore mixed in deionized water. Polymer scaffolds, fabricated from poly(L-lactic-co-glycolic acid) using a solvent casting and particulate leaching technique, are porous enough to absorb serum albumin and can also be doped with various hemostatic and thrombogenic agents to aid in tissue healing. Use of the polymer scaffolds allows one to combine the strength of solid solders and the flexibility of liquid solders without the common “runaway” problems. An in vitro study was performed to correlate tissue temperature with the tensile strength of arterial repairs formed using the chromophore-enhanced solder-doped polymer scaffolds. Laser irradiance was varied and the solder surface and solder/tissue interface temperatures were monitored by an IR temperature monitoring system and a type-K thermocouple, respectively. The solder/tissue interface temperature required for optimized tensile strength was determined to be 67 ± 5°C. This value was in agreement with previous studies using serum albumin solders alone, where the optimal solder/tissue interface temperature was found to be 65°C.
The use of indocyanine green-doped albumin protein solders has been shown to vastly improve the anastomotic strength that can be achieved by laser tissue repair techniques, while at the same time minimizing collateral thermal tissue damage. However, the safety of the degradation products of the chromophore following laser irradiation is uncertain. Therefore, we studied the feasibility of using alternative chromophores in terms of temperature rise at the solder/tissue interface, the extent of thermal damage in the sourrounding tissue, and the tensile strength of repairs. Biodegradable polymer scaffolds of controlled porosity were fabricated with poly(L-lactic-co-glycolic acid), using a solvent-casting and particulate-leaching technique. The porous scaffold acted as a carrier to the traditional protein solder composition of serum albumin and an absorbing chromophore mixed in deionized water. Two commonly used chromophores, indocyanine green and methylene blue were investigated, as well as blue and green food colorings. Temperature rise at the solder surface and at the interface between the solder and tissue were monitored by an IR temperature monitoring system and a type-K thermocouple, respectively, and the extent of thermal damage in the underlying tissue was determined using light microscopy. As expected, temperature rise at the solder/tissue interface, and consequently the degree of collateral thermal tissue damage, was directly related to the penetration depth of the laser light in the protein solder. Optimal tensile strength of repairs was achieved by selecting a chromophore concentration that resulted in a temperature of 66 ± 3°C at the solder/tissue interface.
An investigation was conducted to assess the feasibility ofusing various synthetic polymers as scaffolds to traditional albumin protein solders with the aim of enhancing the mechanical strength of repairs formed during laser tissue soldering. Biodegradable polymer membranes of controlled porosity were fabricated with either polylactic acid (PLA), polyglycolic acid (PGA), or two different poly(L-lactic-co-glycolic acid) (PLGA) blend ratios, using a solvent-casting and particulate-leaching technique. In addition, membranes were prepared by combining each of the above mentioned polymers with poly(ethylene glycol) (PEG). The porous membranes provided a scaffold into which the traditional protein solder composition of serum albumin and indocyanine green dye mixed in deionized water was readily absorbed. Studies were conducted to assess the influence of various processing parameters on the strength of tissue repairs formed using the new adhesives. These parameters included the polymer type, the polymer/PEG blend ratio, the salt particle size, and the initial albumin weight fraction. No significant difference was observed between the four polymers investigated, however, variation of the polymer type and blend ratio is known to influence the degradation rate of the membranes. The addition of PEG to the films during the casting stage was observed to increase the flexibility of the membranes. Finally, repair strength increased significantly with an increase in the size ofpores present in the membranes and with increased protein concentration.
An in vivo study was conducted to investigate the feasibility, mechanical function, and chronic biocompatibility of a new range of light-activated surgical adhesives for vascular anastomosis. Porcine carotid arteries (n=12) and femoral arteries (n=12) were exposed, and a 0.3 -0.6cm longitudinal incision was made in the arterial walls. The vessels were divided equally into two groups. Vessels belonging to the first group were repaired using a surgical adhesive, composed of a poly(L-lactic-co-glycolic acid) scaffold doped with the traditional protein solder mix of serum albumin and indocyanine green dye mixed in deionized water. The adhesive was applied across the incision and denatured using an 805-nm diode laser. Vessels belonging to the second group formed part of a control study, and were repaired using conventional suturing techniques. Blood flow was restored to the vessels immediately after the procedure and the incision sites were checked for patency. The strength and hemostatic abilities of the new surgical adhesives were evaluated in the context of arterial pressure, persistence of hemostasis and presence of any inflammatory reaction after 3 days. The adhesive technique compared favorably with the suture technique. Repairs formed with the adhesive technique were achieved more rapidly than suturing, and acute leakage was observed less frequently. Repairs closed by suture did not burst, but leaked at pressures significantly below those of vessels closed with the adhesive material. Finally, the adhesive technique produced better histology than the suture technique, suggesting that it holds great promise as an alternative to suturing.
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