Low Intensity pulsed Ultrasound an alternative to LSJL

Low Intensity Pulsed Ultrasound has been shown to have profound benefits on the bone.  The theory behind Lateral Synovial Joint Loading is that it works by increasing interstitial fluid flow which in turn increases hydrostatic pressure.  Fluid flow transports stem cells to the hyaline cartilage growth plate line and hydrostatic pressure promotes chondrogenic differentiation of stem cells.  Joint Loading has been shown so far to increase ankle size and there have been sporadic but inconsistent reports of height increase(still self experimenting but need yet more experimenters).  The profound increase in ankle size shows that LSJL may be profoundly effective in increasing height in the spine.  The problem is finding an effective way to load the spine.  Enter LIPUS: Medical Products Online Professional Ultrasound Systems - AC Adapter 240/110 Volt International/Universal Adapter which can be purchased fairly cheaply(the goal is to induce shear strain and induce hydrostatic pressure exact degrees may not matter).

The machine listed above can go up to 150hz.  You can apply the ultrasound device to each of the vertabrae.  Like LSJL, LIPUS may enhance interstitial fluid flow.  So benefits of LSJL and LIPUS may intersect.

Pertussis toxin-sensitive Galphai protein and ERK-dependent pathways mediate ultrasound promotion of osteogenic transcription in human osteoblasts.

"Bone cells respond to mechanical stimulation via mechanoreceptors and convert biophysical stimulation into biochemical signals that alter gene expression and cellular adaptation. Pulsed acoustic energy treatment raises membrane potential and induces osteogenic activity. How membrane-bound osteoblast mechanoreceptors convert physical ultrasound (US) stimuli into osteogenic responses is not fully understood. We demonstrated that low-intensity pulsed US treatment (200-micros pulse, 1 kHz, 30 mW/cm2) elevated Cbfa1/Runx2 mRNA expression and progressively promoted osteocalcin mRNA expression in human osteoblasts. Pretreatment with pertussis toxin (PTX), but not with cholera toxin, suppressed US-augmented osteogenic transcription. This indicated that Gi proteins, but not Gs proteins, were involved in US promotion of osteogenic transcription. Further studies demonstrated US treatment could rapidly increase PTX-sensitive Galphai protein levels and subsequently enhanced phosphorylation of extracellular signal-regulated kinase (ERK). PTX pretreatment significantly reduced US promotion of ERK activation. Moreover, inhibition of ERK activity by PD98059 suppressed US augmentation of Cbfa1/Runx2 and osteocalcin mRNA expression. Membranous Galphai proteins and cytosolic ERK pathways acted as potent mechanosensitive signals in the response of osteoblasts to pulsed US stimulation[so interstitial fluid flow]."

The machine listed above only has a watt output of 8 and the amount tested in this study was 30(since our goal is to stimulate the spine and Runx2 is involved in osteoblasts which increase height in the spine we want to at least match the study).  I don't know where to find a higher powered ultrasound.

Identification of genes responsive to low-intensity pulsed ultrasound stimulations.

"This study was designed to compare the temporal changes of gene expression profile in osteoblastic cell lines (SaOS-2) treated with low-intensity pulsed ultrasound stimulation (LIPUS) using complementary DNA (cDNA) microarrays. SaOS-2 cells were treated with LIPUS for 20min. Thereafter, cells were harvested and RNA was extracted twice at 4 and 24h, respectively. Using cDNA microarrays, 7488 genes with changes in expression in SaOS-2 cells were identified for comparison. Microarray analysis revealed a total of 165 genes in SaOS-2 cells were regulated at 4 and 24h after LIPUS treatment. Except for 30 known LIPUS-regulated genes, our study demonstrated for the first time that over 100 genes were related to the underlying molecular mechanism of LIPUS and suggested that LIPUS might regulate a transient expression of numerous critical genes in osteoblastic cells."

Although this study relates to an osteoblastic cell line the same sort of genes may be upregulated in MSCs and chondrocytes.  And in the future we can use this study to compare to other studies to see which genes are upregulated to osteoblasts and which are upregulated due to other factors.

"(LIPUS, 200-μs burst of 1.5 MHz and 30.0 ± 5.0 mW/cm2) was able to stimulate osteogenesis"

" LIPUS induced transient expression of the immediate-early response gene c-fos and elevated gene expression for bone sialoprotein (BSP), insulin-like growth factor-1 (IGF-1), and osteocalcin (OC) using a mouse bone marrow stromal-derived cell line (ST2 cell), osteoblasts, and osteocytes"

"he LIPUS system used for this study generated 1.5 MHz ultrasound in a pulsed-wave mode (200-μs pulse burst-width with repetition frequency of 1 kHz at the spatial-averaged temporal-averaged intensity of 30 mW/cm2) "

Genes upregulated by LIPUS(noted if only upregulated at 4 or 24 hours) also by LSJL:
ITGBL1
IGFBP6(24hrs only)
GNAL(24hrs only){down}
LOX(24hrs only)

Downregulated:


Early gene response to low-intensity pulsed ultrasound in rat osteoblastic cells

"The aim of the current research was to quantify the changes in gene expression in rat bone marrow derived stromal cells (BMSC) to low intensity pulsed ultrasound (LIPUS) during early time points after the ultrasound application. LIPUS at 1.5 MHz, 30 mW/cm2 was applied to BMSC for a single 20 min treatment. Real-time PCR was carried out to quantify the expression of early response genes and bone differentiation marker genes 0.5, 1, 3, 6 and 12 h after the end of the LIPUS treatment. Compared with the controls, LIPUS treatment resulted in elevated transient expression of early response genes (c-jun{up}, c-myc, COX-2{up as PTGS2}, Egr-1{up}, TSC-22) as well as the bone differentiation marker genes, osteonectin and osteopontin{up}, at 3 h."

At 1 hour in response to LIPUS egr-1, c-jun, and PTGS2 were downregulated in contrast to LSJL where these genes were upregulated after the last loading.  These genes returned to baseline at about 6 hours so it's not likely that the upregulation in LSJL was due to residual effects.

Similar trends occurred with osteopontin and that was measured with RT-PCR whereas the responses to the other three genes were done with microarray for LSJL.