Second harmonic generation microscopy of fetal membranes under deformation: normal and altered morphology.
BACKGROUND
Insight into the microstructure of fetal membrane and its response to deformation is important for understanding causes of preterm premature rupture of the membrane. However, the microstructure of fetal membranes under deformation has not been visualized yet. Second harmonic generation microscopy, combined with an in-situ stretching device, can provide this valuable information.


METHODS
Eight fetal membranes were marked over the cervix with methylene blue during elective caesarean section. One sample per membrane of reflected tissue, between the placenta and the cervical region, was cyclically stretched with a custom built inflation device. Samples were mounted on an in-situ stretching device and imaged with a multiphoton microscope at different deformation levels. Microstructural parameters such as thickness and collagen orientation were determined. Image entropy was evaluated for the spongy layer.


RESULTS
The spongy layer consistently shows an altered collagen structure in the cervical and cycled tissue compared with the reflected membrane, corresponding to a significantly higher image entropy. An increased thickness of collagenous layers was found in cervical and stretched samples in comparison to the reflected tissue. Significant collagen fibre alignment was found to occur already at moderate deformation in all samples.


CONCLUSIONS
For the first time, second harmonic generation microscopy has been used to visualize the microstructure of fetal membranes. Repeated mechanical loading was shown to affect the integrity of the amnion-chorion interface which might indicate an increased risk of premature rupture of fetal membrane. Moreover, mechanical loading might contribute to morphological alterations of the fetal membrane over the cervical region.
Department of Mechanical and Process Engineering, ETH Zurich, 8092 Zurich, Switzerland. Electronic address: mauri@imes.mavt.ethz.ch.
A
A
Mauri
M
M
Perrini
J M
JM
Mateos
C
C
Maake
N
N
Ochsenbein-Koelble
R
R
Zimmermann
M
M
Ehrbar
E
E
Mazza
D002584:Cervix Uteri; D002585:Cesarean Section; D055598:Chemical Phenomena; D005109:Extracellular Matrix / Q000737:chemistry / Q000473:pathology*; D005321:Extraembryonic Membranes / Q000737:chemistry / Q000166:cytology / Q000473:pathology*; D005260:Female; D005322:Premature Rupture of Fetal Membranes / Q000453:epidemiology / Q000473:pathology*; D024022:Fibrillar Collagens / Q000737:chemistry; D006801:Humans; D066298:In Vitro Techniques; D055595:Mechanical Phenomena; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D036641:Microscopy, Fluorescence, Multiphoton; D008954:Models, Biological; D009215:Myometrium; D009929:Organ Size; D011247:Pregnancy; D012306:Risk; D013314:Stress, Mechanical; D013557:Switzerland / Q000453:epidemiology; D016474:Weight-Bearing
D024022:Fibrillar Collagens
Fetal membrane; Multiphoton microscopy; Second harmonic generation microscopy; Zone of altered morphology