Formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium

Background: The bioluminescent enzyme firefly luciferase (Luc) or variants of green fluorescent protein (GFP) in transformed cells can be effectively used to reveal molecular and cellular features of neoplasia in vivo. Tumor cell growth and regression in response to various therapies can be evaluated by using bioluminescent imaging. In bioluminescent imaging, light propagates in highly scattering tissue, and the diffusion approximation is sufficiently accurate to predict the imaging signal around the biological tissue. The numerical solutions to the diffusion equation take large amounts of computational time, and the studies for its analytic solutions have attracted more attention in biomedical engineering applications. Methods: Biological tissue is a turbid medium that both scatters and absorbs photons. An accurate model for the propagation of photons through tissue can be adopted from transport theory, and its diffusion approximation is applied to predict the imaging signal around the biological tissue. The solution to the diffusion equation is formulated by the convolution between its Green's function and source term. The formulation of photon diffusion from spherical bioluminescent sources in an infinite homogeneous medium can be obtained to accelerate the forward simulation of bioluminescent phenomena. Results: The closed form solutions have been derived for the time-dependent diffusion equation and the steady-state diffusion equation with solid and hollow spherical sources in a homogeneous medium, respectively. Meanwhile, the relationship between solutions with a solid sphere source and ones with a surface sphere source is obtained. Conclusion: We have formulated solutions for the diffusion equation with solid and hollow spherical sources in an infinite homogeneous medium. These solutions have been verified by Monte Carlo simulation for use in biomedical optical imaging studies. The closed form solution is highly accurate and more computationally efficient in biomedical engineering applications. By using our analytic solutions for spherical sources, we can better predict bioluminescent signals and better understand both the potential for, and the limitations of, bioluminescent tomography in an idealized case. The formulas are particularly valuable for furthering the development of bioluminescent tomography

A Born-type approximation method for bioluminescence tomography

In this paper, we present a Born-type approximation method for bioluminescence tomography (BLT), which is to reconstruct an internal bioluminescent source from the measured bioluminescent signal on the external surface of a small animal. Based on the diffusion approximation for the photon propagation in biological tissue, this BLT method utilizes the Green function to establish a linear relationship between the measured bioluminescent signal and the internal bioluminescent source distribution. The Green function can be modified to describe a heterogeneous medium with an arbitrary boundary using the Born approximation. The BLT reconstruction is formulated in a linear least-squares optimization framework with simple bounds constraint. The performance of this method is evaluated in numerical simulation and phantom experiments

Development of lung tissue phantoms for bioluminescent imaging

White nylon material was chosen to make cylindrical tissue phantoms for development of bioluminescence tomography techniques. A low-level light source, delivered through an optic fiber of core diameter 200 μm, was placed at different locations on one phantom surface. The light travels through the phantom, reaches the external surface, and is captured by a liquid nitrogen-cooled CCD camera. The scattering, absorption, and anisotropy parameters of the phantom are obtained by matching the measured light transmission profiles to the profiles generated by the TracePro software. The perturbation analysis, with the homogeneous phantoms, demonstrated that the imaging system is sufficiently sensitive to capture intensity change of higher than 0.5nW/cm2 or a location shift of the light source of more than 200 microns. It is observed that the system can distinguish two point light sources with separation of about 2 mm. The perturbation analysis is also performed with the heterogeneous phantom. Based on our data, we conclude that there is inherent tomographic information in bioluminescent measures taken on the external surface of the mouse, which suggests the feasibility of bioluminescence tomography for biomedical research using the small animals, especially the mice

Generation of a recombinant rabies Flury LEP virus carrying an additional G gene creates an improved seed virus for inactivated vaccine production

The rabies Flury Low Egg Passage virus (LEP) has been widely used as a seed virus to generate inactive vaccine. Here, we established a reverse genetic system for LEP and generated a recombinant LEP virus (rLEP-G) that carries two identical G genes. This recombinant virus showed similar properties to those of LEP with respect to in vitro growth, neurotropism index, and virulence in mice. rLEP-G produced 4.3-fold more G protein than did LEP in BHK-21 cells. The inactivated vaccine generated from rLEP-G induced significantly higher virus neutralization titers in mice and dogs than those produced in response to LEP-derived vaccine. Our results suggest that rLEP-G is an improved seed virus candidate for inactivated rabies virus vaccine manufacture

In vivo tomographic imaging based on bioluminescence

The most important task for bioluminescence imaging is to identify the emission source from the captured bioluminescent signal on the surface of a small tested animal. Quantitative information on the source location, geometry and intensity serves for in-vivo monitoring of infectious diseases, tumor growth, metastases in the small animal. In this paper, we present a point-spread function-based method for reconstructing the internal bioluminescent source from the surface light output flux signal. The method is evaluated for sensing the internal emission sources in nylon phantoms and within a live mouse. The surface bioluminescent signal is taken with a highly sensitive CCD camera. The results show the feasibility and efficiency of the proposed point-spread function-based method

Development of lung tissue phantoms for bioluminescent imaging

White nylon material was chosen to make cylindrical tissue phantoms for development of bioluminescence tomography techniques. A low-level light source, delivered through an optic fiber of core diameter 200 μm, was placed at different locations on one phantom surface. The light travels through the phantom, reaches the external surface, and is captured by a liquid nitrogen-cooled CCD camera. The scattering, absorption, and anisotropy parameters of the phantom are obtained by matching the measured light transmission profiles to the profiles generated by the TracePro software. The perturbation analysis, with the homogeneous phantoms, demonstrated that the imaging system is sufficiently sensitive to capture intensity change of higher than 0.5nW/cm2 or a location shift of the light source of more than 200 microns. It is observed that the system can distinguish two point light sources with separation of about 2 mm. The perturbation analysis is also performed with the heterogeneous phantom. Based on our data, we conclude that there is inherent tomographic information in bioluminescent measures taken on the external surface of the mouse, which suggests the feasibility of bioluminescence tomography for biomedical research using the small animals, especially the mice

Mutant GDF5 enhances ameloblast differentiation via accelerated BMP2-induced Smad1/5/8 phosphorylation

Bone morphogenetic proteins (BMPs) regulate hard tissue formation, including bone and tooth. Growth differentiation factor 5 (GDF5), a known BMP, is expressed in cartilage and regulates chondrogenesis, and mutations have been shown to cause osteoarthritis. Notably, GDF5 is also expressed in periodontal ligament tissue; however, its role during tooth development is unclear. Here, we used cell culture and in vivo analyses to determine the role of GDF5 during tooth development. GDF5 and its associated BMP receptors are expressed at the protein and mRNA levels during postnatal tooth development, particularly at a stage associated with enamel formation. Furthermore, whereas BMP2 was observed to induce evidently the differentiation of enamel-forming ameloblasts, excess GDF5 induce mildly this differentiation. A mouse model harbouring a mutation in GDF5 (W408R) showed enhanced enamel formation in both the incisors and molars, but not in the tooth roots. Overexpression of the W408R GDF5 mutant protein was shown to induce BMP2-mediated mRNA expression of enamel matrix proteins and downstream phosphorylation of Smad1/5/8. These results suggest that mutant GDF5 enhances ameloblast differentiation via accelerated BMP2-signalling