Denaturation of collagen via heating: an irreversible rate process.

Heating therapies are increasingly used in cardiology, dermatology, gynecology, neurosurgery, oncology, ophthalmology, orthopedics, and urology, among other medical specialties. This widespread use of heating is driven primarily by the availability of new technology, not by a detailed understanding of the biothermomechanics. Without basic quantification of the underlying physical and chemical processes in terms of parameters that can be controlled clinically, identification of preferred interventions will continue to be based primarily on trial and error, thus necessitating large clinical studies and years of accumulative experience. Perusal of the literature reveals that much has been learned over the past century about the response of cells, proteins, and tissues to supra-physiologic temperatures; yet, the associated findings are reported in diverse journals and the underlying basic processes remain unidentified. In this review, we seek to contrast various findings on the kinetics of the thermal denaturation of collagen and to encourage investigators to consider the many open problems in part via a synthesis of results from the diverse literatures.

Vaccinia virus-free recovery of vesicular stomatitis virus.

The advent of reverse-genetics represents a powerful new approach to elucidate aspects of negative-sense RNA virus replication. The reverse-genetics system established previously for vesicular stomatitis virus (VSV) required four plasmids encoding the nucleoprotein (N), phosphoprotein (P), polymerase (L), and the full-length, anti-genomic RNA. Transcription to yield the antigenomic RNA as well as the N, P, and L, mRNAs was initiated by bacteriophage T7 polymerase expressed from a recombinant Vaccinia virus. In this report, we describe the successful recovery of infectious VSV in the absence of Vaccinia virus. The N, P, and L genes of VSV were inserted downstream of both the T7 promoter and an internal ribosomal entry site (IRES element). T7 polymerase was expressed constitutively from BSR-T7/5 cells. RTPCR was used to confirm that the recovered VSV was derived from transfected DNA. Virion protein profile, CPE in tissue culture, and virus titer of the recombinant VSV were indistinguishable from those of parental VSV. Thus, the need for Vaccinia virus is eliminated with this system, making it an attractive, alternative approach for the recovery of infectious VSV from DNA.

Effects of heat-induced damage on the radial component of thermal diffusivity of bovine aorta.

The extent of the change in thermal diffusivity of soft tissues due to heat-induced damage is not well known. Reported here are the results of using the flash method to measure the through-the-wall component of thermal diffusivity of bovine aorta before and after the tissue has undergone two hours of heating at 75 degrees C. The measurements indicate a 10.1 percent increase in the thermal diffusivity of the tissue post-heating. While this change may not result in a significant change in the tissue temperature profile, further study is needed to quantify the thermal diffusivity in other coordinate directions, as well as the mechanisms by which this change in properties occurs.

Temperature elevation caused by bone cement polymerization during vertebroplasty.

Percutaneous vertebroplasty (PVP), whereby polymethylmethacrylate cement is injected into the vertebral body (VB), has been used to successfully treat various spinal lesions. The mechanism responsible for the palliative effect of PVP is unknown, but it may be the result of neural damage caused by heat liberated during polymerization of the polymethylmethacrylate. The purpose of the current study was to measure in vitro temperature histories at three key locations (anterior cortex, center, spinal canal) in VBs injected with one of two different bone cements (Simplex P and Orthocomp) to determine the role temperature plays in PVP. Twelve VBs (T11-L2) from three elderly female spines were instrumented with thermocouples and injected with 10 cc of one of the two cements. Temperatures were measured with the VBs in a bath (37 degrees C) for 15 min after injection. A Student's paired t-test was used to determine differences in peak temperature and time above 50 degrees C between the two cement groups. Peak temperatures and temperatures above 50 degrees C were significantly higher and longer, respectively, at the center of VBs injected with Simplex P (61.8 +/- 12.7 degrees C; 3.6 +/- 2.1 min) than those injected with Orthocomp (51.2 +/- 6.2 degrees C; 1.3 +/- 1.4 min). There was no significant difference in peak temperature between cements at the spinal canal location; temperature there did not rise above 41 degrees C. Although thermal damage to intraosseous neural tissue caused by cement polymerization cannot be ruled out as a potential mechanism for pain relief experienced by patients subsequent to PVP, it seems unlikely based on the worst-case conditions tested in the current study.

Phenomenological evolution equations for heat-induced shrinkage of a collagenous tissue.

Optimization of clinical heat treatments for various pathologies requires accurate numerical modeling of the heat transfer, evolution of thermal damage, and associated changes in the material properties of the tissues. This paper presents two phenomenological equations that quantify time-dependent thermal damage in a uniaxial collagenous tissue. Specifically, an empirical rule-of-mixtures model is shown to describe well heat-induced axial shrinkage (a measure of underlying denaturation) in chordae tendineae which results from a spectrum of thermomechanical loading histories. Likewise an exponential decay model is shown to describe well the partial recovery (e.g., renaturation) of chordae when it is returned to body temperature following heating. Together these models provide the first quantitative descriptors of the evolution of heat-induced damage and subsequent recovery in collagen. Such descriptors are fundamental to numerical analyses of many heat treatments because of the prevalence of collagen in many tissues and organs.

Measurement of thermal diffusivity of bovine aorta subject to finite deformation.

The flash thermal diffusivity measurement technique is applied to tissue for the first time. Making use of its minimal contact with the specimen, the flash technique is extended to allow for well-defined, biaxial, finite strain. As an example application, the radial component of thermal diffusivity of bovine descending aorta is measured in vitro as a function of equibiaxial stretch, at room temperature. Data analysis is accomplished using a Marquardt algorithm coupled with a finite difference solution of the thermal diffusion equation. Extension of this method to measure simultaneously three orthogonal components of diffusivity, at different levels of temperature, is discussed.

Heat-induced changes in the mechanics of a collagenous tissue: isothermal, isotonic shrinkage.

We present data from isothermal, isotonic-shrinkage tests wherein bovine chordae tendineae were subjected to well-defined constant temperatures (from 65 to 90 degrees C), durations of heating (from 180 to 3600 s), and isotonic uniaxial stresses during heating (from 100 to 650 kPa). Tissue response during heating and "recovery" at 37 degrees C following heating was evaluated in terms of the axial shrinkage, a gross indicator of underlying heat-induced denaturation. There were three key findings. First, scaling the heating time via temperature and load-dependent characteristic times for the denaturation process collapsed all shrinkage data to a single curve, and thereby revealed a time-temperature-load equivalency. Second, the characteristic times exhibited an Arrhenius-type behavior with temperature wherein the slopes were nearly independent of applied load--this suggested that applied loads during heating affect the activation entropy, not energy. Third, all specimens exhibited a time-dependent, partial recovery when returned to 37 degrees C following heating, but the degree of recovery decreased with increases in the load imposed during heating. These new findings on heat-induced changes in tissue behavior will aid in the design of improved clinical heating protocols and provide guidance for the requisite constitutive formulations.

Heat-induced changes in the mechanics of a collagenous tissue: isothermal free shrinkage.

We present data from isothermal free-shrinkage tests (i.e., performed in the absence of mechanical loads) wherein bovine chordae tendineae were subjected to temperatures from 65 to 85 degrees C for 120 to 1200 s. These data reveal four new insights into heat-induced denaturation of a collagenous tissue. First, a characteristic time for the free shrinkage appears to exhibit an Arrhenius-type relationship with temperature. Second, scaling the actual heating time via the characteristic time results in a single correlation between free shrinkage and the duration of heating; this correlation suggests a time-temperature equivalence. Third, it is the cumulative, not current, heating time that governs the free shrinkage. And fourth, heat-induced free shrinkage is partially recovered when the tissue is returned to 37 degrees C, this recovery also being time-dependent. Although these findings will help guide future experimentation and constitutive modeling, as well as the design of new heat-based clinical therapies, there is a pressing need to collect additional isothermal data, particularly in the presence of well-defined mechanical loads.